Search

Fil: Femia, Lis. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Desarrollo Tecnologico para la Industria Quimica. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnologico para la Industria Quimica; Argentina

The controlled synthesis of polymer nanoparticles is of great interest in the production of latexes with well defined characteristics. These products can be applied in biomedicine as carriers of biomolecules (e.g. proteins and enzymes), and in particular as latexes for immunoassays used for example in immunoagglutination test, which allow amplify the antigen–antibody reaction, being simple, quick and inexpensive diagnostic tools. The synthesis of polystyrene (PS) latex particles and of core–shell particles, with controlled size distribution, functional groups and surface charge densities is considered here. PS latexes were synthesized in the 100–1100 nm diameter range, by either emulsion or dispersion polymerization. Such latexes were then used as seeds in emulsion copolymerizations of styrene and a functional monomer (methacrylic acid or glycidyl methacrylate), thus producing particles with carboxyl or epoxy functionalities, respectively. Changes in the polymerization recipes employed under batch operation were analyzed. Latex characterization involved measurements of mean particle diameters, the polydispersity index, functional group densities and zeta potential. Finally, latexes were sensitized (by either physical adsorption or covalent coupling) with specific antigens to obtain latex–protein complexes, and one of them was tested as agglutination assay for detecting leptospirosis disease in bovine samples, as an example of the potential application of the latexes produced. Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Clementi, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

Nanogels play a leading role in controlled release systems because they possess high water retention capacity resulting in high loading capabilities, stability in biological fluids and biocompatibility. In this scenario, every tool that allows extending the nanogel properties and expanding their potential applications is of high interest in the field of biomedicine. This article aims to contribute to the development of multifunctional nanogels, based on the combination of two polymer phases in a multilobular morphology. The synthesized multilobed nanogels (mLNGs) presented a core of crosslinked poly(N-vinylcaprolactam) (PVCL) and a shell formed by 3-D distributed lobes of a low Tg copolymer. This particular multilobular morphology is able to exploit the synergetic contribution of both phases. While the PVCL-based core conferred its characteristic thermal response and the ability to load and release a cargo molecule, the low Tg lobes incorporated the capability of film formation. Moreover, the multilobular arrangement of NGs allows films to undergo unrestricted mass transfer. The development of mLNG morphology and the effect of synthesis parameters were deeply studied with the help of a previously developed mathematical model for the dynamic evolution of particle morphology. Finally, this study presents, for the first time, the synthesis of two-phase nanogels with multilobular morphology and underlines their potential as a candidate for controlled delivery platforms.

Latex-particles are suitable for diagnosis purpose since they are able to interact with biological molecules. In this work, a colored monomer was synthesized between basic fuchsine (BF) and glycidyl methacrylate (GMA). BF-GMA obtaining was verified through UV-Vis, IR, NMR and Mass spectrometry. The synthesis of colored particles by emulsion polymerization of styrene and the BF-GMA monomer was studied by measuring conversion, particle diameter, BF incorporation, surface charge densities and also in a protein binding assay. Results confirmed the production of stable particles, with controlled size and high BF incorporation, capable of interacting with proteins efficiently, and being used in an immunoagglutination test. Fil: Femia, Lis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

The aim of the present work was to synthesize sustained-release hydrogels based on N,N-dimethylacrylamide (DMA) to study the effect of the polymer matrix structure and the solubility of drugs on the load and release mechanisms. A series of cross-linked DMA hydrogels were synthesized with different monomer concentration and crosslinker to monomer ratio by free radical aqueous solution polymerization at 37ºC using N,N-methylen-bis-acrylamide as crosslinking agent. The effect of total monomer concentration and degree of crosslinking on the water absorption, glass transition temperature values, mechanical properties, and load/release of different drugs was studied. Main results showed that: a) the structure of the network, obtained by varying the monomer and crosslinking agent concentration, is a factor that governs the chemical and physical properties of the hydrogel (water absorption, glass transition temperature, storage and loss moduli, network parameters), b) the rate and amount of drug released from swellable hydrogels depend on both the degree of hydrogel crosslinking and the water solubility of the drug. In all cases, as the concentration of the crosslinker agent increased, the swelling capacity of the hydrogels was reduced and drugs with high solubility in water were more easily released, probably due to their greater affinity for the medium. Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

The chemical coupling of a protoplasmatic antigen from Mycobacterium avium subsp. paratubeculosis onto core-shell carboxylated particles was investigated with the aim of producing latex-protein complexes to be used in immunoagglutination assays capable of detecting bovine paratuberculosis disease. For this purpose, sensitizations were carried out using both colored and not colored carboxylated latexes as well as the protoplasmatic antigen at pH close to its isoelectric point to favor the antigenic protein to approach the particle surface. In all cases, higher fractions of proteins were chemically-bound to carboxyl groups on the surface of the particles. The assessment of the performance of the visual immunoagglutination assays consisted of evaluating 111 sera from healthy and infected bovines with Mycobacterium avium subsp. paratuberculosis. Complexes obtained from the colored latex allowed an acceptable visual discrimination between the studied positive and negative sera. Most of the positive samples showed strong to very strong agglutination and only a few samples reacted weakly, i.e. a sensitivity of 70%. The specificity of the assay, on the other hand, was 86%. Therefore, this rapid detection technique allows an easy and inexpensive identification of animals possibly infected with paratuberculosis "in situ" in the herds.

The daily clinical practice frequently force physicians to make adecision of whether a patient is coursing an infectious disease at theacute or the chronic stage, since treatment may be different dependingon it. This is the case of toxoplasmosis, a worldwide disease caused bythe intracellular parasite Toxoplasma gondii. When a pregnant woman isinfected for the first time by T. gondii and is not treated, the parasite cancross the placenta and severely affect the fetus. In case of primary infection,the treatment protocol includes using antiparasitic drugs toprevent serious damages, including death of the unborn baby. However,antiparasitic chemicals have side effects that should be avoided, unlesstreatment is mandatory. That is why discrimination between long-termand primary T. gondii infection turns out to be critical in infectedpregnant women (Montoya and Remington, 2008).In this work we have used rP22a, an acute-phase recombinant protein werecently described (Costa et al., 2017), to assess anti-T. gondii IgG AI,calculated by two new easier approaches, one based on the area underthe avidity curve (AUAC) and the other one based on the E-P titrationmethod with minor variations. We compare our results with those obtainedwhen calculating the AI by the conventional methods abovementioned, and the performance to render a proper sample classificationas compared to that obtained having used a commercial kit asstandard. Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Peverengo, Luz María. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Dalla Fontana, Maria L.. Laboratorio Central de la Provincia de Santa Fe; Argentina Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lagier, Claudia Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina

Transdermal immunization is highly attractive because of the skin's accessibility and unique immunological characteristics. However, it remains a relatively unexplored route of administration because of the great difficulty of transporting antigens past the outermost layer of skin, the stratum corneum. In this article, the abilities of three poly(N-vinylcaprolactam) (PVCL)-based thermoresponsive assemblies - PVCL hydrogels and nanogels plus novel film forming PVCL/acrylic nanogels - to act as protein delivery systems were investigated. Similar thermal responses were observed in all systems, with transition temperatures close to 32 °C, close to that of the skin surface. The investigated dermal delivery systems showed no evidence of cytotoxicity in human fibroblasts and were able to load and release ovalbumin (OVA), a well-studied antigen, in a temperature-dependent manner in vitro. The penetration of OVA into ex vivo human skin following topical application was evaluated, where enhanced skin delivery was seen for the OVA-loaded PVCL systems relative to administration of the protein alone. The distinct protein release and skin penetration profiles observed for the different PVCL assemblies were here discussed on the basis of their structural differences. Fil: Sonzogni, Ana Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Yealland, Guy. Universidad Libre de Berlin; Alemania Fil: Kar, Mrityunjoy. Universidad Libre de Berlin; Alemania Fil: Wedepohl, Stefanie. Universidad Libre de Berlin; Alemania Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Hedtrich, Sarah. Universidad Libre de Berlin; Alemania Fil: Calderon, Marcelo. Universidad Libre de Berlin; Alemania Fil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

Canine visceral leishmaniasis (CVL) is the major source of human visceral leishmaniasis. To control the spread of this disease, early and accurate detection of infected dogs is critical but challenging. The serological diagnosis of CVL remains problematic because there are no reliable commercially available tests. Most laboratories use enzyme-linked immunosorbent assay or the indirect immunofluorescent antibody test. These tests use Leishmania chagasi recombinant antigens K39 or K26 assembled with either gold-labelled Staphylococcus aureus protein A or protein G from Streptococcus pyogenes. In this work, we propose the development, optimization and standardization of a lateral flow immunoassay (LFIA) based on functionalized colored particles and a specific recombinant antigen, as a visual in situ method for the diagnosis of CVL. The following analysis variables were considered: i) the concentration of the latex-protein complex; ii) the dilution of the serum; iii) the composition of the employed buffers; iv) the nominal capillary flow time through the nitrocellulose membrane; v) the concentration of reagents fixed in the test and control lines; vi) the particle size of the colored latex; and vii) the conjugation method. Then, the obtained strips were evaluated as a visual diagnostic tool based on a panel of positive and negative sera. It was observed that because of its simplicity and performance the LFIA test is a quick and reliable alternative for the diagnosis of CVL either in conventional laboratories or for remote areas where laboratories are not readily accessible for conventional assays. Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Guerrero, Sergio Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina. Universidad Nacional del Litoral. Facultad de Ingeniería Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina

Nowadays material science is trying to design products that combine the adaptability of synthetic polymers with the structure and functionality of biopolymers. In this context, the present work investigates the production of hybrid nanocomposites based on acrylic and hydrolyzed collagen (HC) to obtain a waterborne dispersion with film forming capability. For this purpose, the emulsion copolymerization of acrylic acid (AA) and butyl acrylate (BA) in the presence of different concentrations of HC was investigated. The synthesized acrylic?HC latexes are able to form films with promising properties for their potential application as bio-adhesives, where adhesiveness could be controlled by both the degree of neutralization and the moisture content. Fil: Luque, Gisela Carina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Stürtz, Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Passeggi, Mario Cesar Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Física del Litoral. Universidad Nacional del Litoral. Instituto de Física del Litoral; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

The use of chimeric molecules fusing several antigenic determinants is a promising strategy for the development of low-cost, standardized and reliable kits to determine specific antibodies. In this study, we designed and assessed a novel recombinant chimera that complements the performance of our previously developed chimera, CP1 [FRA and SAPA antigens (Ags)], to diagnose chronic Chagas disease. The new chimeric protein, named CP3, is composed of MAP, TcD and TSSAII/V/VI antigenic determinants. We compared the performance of both chimeric Ags using a panel of 67 Trypanosoma cruzi-reactive sera and 67 non-reactive ones. The sensitivity of CP3 vs CP1 was 100 and 90.2%, and specificity was 92.5 and 100%, respectively. The mixture of CP1 + CP3 achieved 100% of sensitivity and specificity. More importantly, an additional subset of 17 sera from patients with discordant results of conventional serological methods was analysed; the CP1 + CP3 mixture allowed us to accurately classify 14 of them with respect to IIF, the usual technique used in most of the reference centres. These results show an improved performance of the CP1 + CP3 mixture in comparison with enzyme-linked immunosorbent assay and indirect haemagglutination commercial assays.

Thermoresponsive nanogels (NGs) with film-forming ability could have great potential in delivery system platforms with controlled and targeted release of drugs or proteins, such as in topical treatment. This platform offers the opportunity of combining both the high loading capacity of an NG dispersion and the delivery capability from a film, which could be previously obtained or directly formed onto the surface where the release is required. Therefore, this article investigates the synthesis of NGs based on poly(N-vinylcaprolactam), a thermoresponsive polymer, and poly(butyl acrylate), which promotes particle coalescence and cohesivity (i.e., film forming). The synthesized NGs have low cytotoxicity and are able to form flexible and smooth films, conserving the thermoresponsivity of the particulated system. The obtained films loaded with a model protein show delivery profiles responsive to temperature changes. Fil: Sonzogni, Ana Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Passeggi, Mario Cesar Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Física del Litoral. Universidad Nacional del Litoral. Instituto de Física del Litoral; Argentina Fil: Wedepohl, Stefanie. Freie Universität Berlin; Fil: Calderon, Marcelo. Universidad Nacional de Córdoba; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

Objectives: To obtain and assess stable cage-like particles with low surface charge density, which can be prepared using a standardized, economic and scalable method. Methods: To form these nanoparticles, the lipid composition and proportion as well the method were modified in relation to cage-like particles previously described elsewhere. Bovine albumin was used to compare ISPA performance with that of other adjuvants in mice and to assess stability. Adjuvant efficacy was analysed using a mouse model of Trypanosoma cruzi infection, which shows protection against an intracellular infection that needs a strong cellular response. Key findings: The new particles were better in terms of level, kinetics and profile of humoral responses than Freund Adjuvant, aluminium hydroxide and Montanide TM ISA 206; they also tended to improve ISCOMATRIX? performance. Particle size and adjuvant performance were conserved during the 6-month period assessed after preparation. In the model of Trypanosoma cruzi infection, mice immunized with ISPA and trans-sialidase developed high protection. Conclusions: The obtained nanoparticles were stable and outperformed the other assessed adjuvants in joining together the capacity of most adjuvants to enhance the immune response against specific antigen, to reduce the number of doses, to homogenize the response between individuals and to reach a balanced TH1/TH2 response. Fil: Bertona, Daiana. Universidad Nacional del Litoral; Argentina Fil: Pujato, Nazarena. Universidad Nacional del Litoral; Argentina Fil: Bontempi, Iván. Universidad Nacional del Litoral; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina. Universidad Nacional del Litoral; Argentina Fil: Cabrera, Gabriel Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral; Argentina Fil: Gugliotta, Luis Marcelino. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Hozbor, Daniela Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Nicastro, Alcides. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Calvinho, Luis Fernando. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral; Argentina

Visceral leishmaniasis is a public health problem worldwide. The early diagnosis in dogs is crucial, since they are an epidemiologically relevant reservoir of the disease. The aim of a field study is to early identify the disease allowing rapid intervention to reduce its effects. We propose an immunoagglutination test as a visual in situ method for diagnosis of canine visceral leishmaniasis. Latex-protein complexes were sensitized by covalent coupling of a chimeric recombinant antigen of Leishmania spp. onto polystyrene latex with carboxyl functionality. The reaction time and the antigen concentration under which the immunoagglutination assay shows greater discrimination between the responses of a positive control serum and a negative control serum were determined. Then, the latex-protein complexes were evaluated as a visual diagnostic tool with a panel of 170 sera. The test may be read between 2 and 5 min and can be performed even using sera with elevated concentration of lipids, bilirubin or with variable percentage of hemolysis. The sensitivity, the specificity and the diagnostic accuracy were 78%; 100% and >80%, respectively. The visual immunoagglutination test is of potential application as a method for field studies because it shows results in less than 5 min, it is easy to implement and does not require sophisticated equipment. Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Burna, Alexis Norberto. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina Fil: Demonte, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Arias, Diego Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Cabeza, Matías Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Guerrero, Sergio Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina

Hybrid nanoparticles containing proteins have a technological interest because they attempt to achieve improved properties with respect to the single materials by chemically linking both components. In this article, the batch emulsion polymerization of methyl methacrylate in the presence of varied concentration of casein and tert-butyl hydroperoxide as initiator was investigated. A detailed characterization of the molecular microstructure and morphology of the hybrid nanoparticles allowed the identification of two competitive particle formation mechanisms. Compatibilized nanoparticles were produced at the beginning of the polymerization, while uncompatibilized particles could be generated by a second way of nucleation, which is promoted by the ungrafted protein and depends on its concentration. Fil: Picchio, Matías Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Passeggi, Mario Cesar Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Investigaciones en Tecnología Química; Argentina Fil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Barandiaran, María. Universidad del Pais Vasco; España Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina

The synthesis and characterization of latex-protein complexes (LPC), from the acute phase recombinant antigen P35 (P35Ag) of Toxoplasma gondii and "core-shell" carboxylated or polystyrene (PS) latexes (of different sizes and charge densities) is considered, with the aim of producing immunoagglutination reagents able to detect recently acquired Toxoplasmosis. Physical adsorption (PA) and chemical coupling (CC) of P35Ag onto latex particles at different pH were investigated. Greater amounts of adsorbed protein were obtained on PS latexes than on carboxylated latexes, indicating that hydrophobic forces govern the interactions between the protein and the particle surface. In the CC experiments, the highest amount of bound protein was obtained at pH 6, near the isoelectric point of the protein (IP = 6.27). At this pH, it decreased both the repulsion between particle surface and protein, and the repulsion between neighboring molecules. The LPC were characterized and the antigenicity of the P35Ag protein coupled on the particles surface was evaluated by Enzyme-Linked ImmunoSorbent Assay (ELISA). Results from ELISA showed that the P35Ag coupled to the latex particles surface was not affected during the particles sensitization by PA and CC and the produced LPC were able to recognize specific anti-P35Ag antibodies present in the acute phase of the disease. Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina Fil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina

Objective To determine the conditions under which the immunoagglutination assay to detect Chagas disease, obtained from a novel latex-(chimeric recombinant antigen) complex, shows greater discrimination between the responses of a positive control serum and a negative control serum. Methods The following variables were determined: (i) the sensitisation mechanism, (ii) the emulsifier employed for protein desorption, (iii) the reaction time, (iv) the ionic strength of the reaction medium, (v) the particle concentration, (vi) the presence of blocking agents, (vii) the presence of polyethyleneglycol as potentiator of reaction and (viii) the antigen and antibody concentrations. The search of optimal conditions was investigated by varying one variable at a time. To this effect, monodisperse latex particles sensitised with a recombinant chimeric protein (CP1) were subjected to different conditions. The agglutination reaction was followed by measuring the changes in the optical absorbance by turbidimetry. Results The maximum discrimination between negative and positive sera was obtained at a reaction time of 5 min, when latex complexes with a concentration of covalently coupled protein of 2.90 mg/m2 were put in contact with undiluted sera in buffer borate pH 8–20 mm containing glycine (0.1 m) and polyethyleneglycol 8000 (3% w/v). Finally, the latex–protein complex was tested under the obtained optimal conditions, with a panel of Trypanosoma cruzi-positive sera, leishmaniasis-positive sera and -negative sera for both parasites. Conclusion The immunoagglutination test based on the latex-CP1 complex could be used as a screening method for detecting Chagas disease. This test is rapid, easy to implement and could be used under field conditions; but its results should be confirmed by reference techniques like ELISA, HAI, and IFI. Objectif Determiner les conditions sous lesquelles le test d'immunoagglutination pour la detection de la maladie de Chagas, obtenu a partir d'un nouveau complexe latex-(antigene recombinant chimerique), montre une plus grande discrimination entre les reponses d'un serum controle positif et d'un controle negatif. Methodes Les variables suivantes ont ete determinees: (i) le mecanisme de sensibilisation, (ii) l’emulsifiant utilise pour la proteine de desorption, (iii) le temps de reaction, (iv) la force ionique du milieu reactionnel, (v) la concentration de particules, (vi) la presence d'agents bloquant, (vii) la presence de polyethylene glycol comme potentialisateur de la reaction et (viii) les concentrations d'antigene et d'anticorps. La recherche des conditions optimales a ete effectuee en faisant varier une variable a la fois. A cet effet, des particules de latex mono disperses, sensibilisees avec une proteine chimerique recombinante (CP1) ont ete soumises a differentes conditions. La reaction d'agglutination a ete suivie par la mesure des variations de l'absorbance optique par turbidimetrie. Resultats La discrimination maximale entre les serums negatifs et positifs a ete obtenue a un temps de reaction de 5 min, lorsque les complexes latex a une concentration de proteine couplee de maniere covalente de 2.90 mg/m2 ont ete mis en contact avec le serum non dilue dans un tampon borate pH 8 de 20 mm contenant de la glycine (0.1 m) et du polyethylene glycol 8000 (3% v/v). Enfin, le complexe latex-proteine a ete teste dans les conditions optimales obtenues, avec une serie de sera positifs pour T. cruzi et la leishmaniose et de sera negatifs pour les deux parasites. Conclusion Le test d'immunoagglutination base sur le complexe latex-CP1 pourrait etre utilise comme une methode de depistage permettant de detecter la maladie de Chagas. Ce test est rapide, facile a implementer et pourrait etre utilise dans des conditions de terrain, mais ses resultats devraient etre confirmes par des techniques de reference comme ELISA, HAI et IFI. Objetivo Determinar las condiciones bajo las cuales el ensayo de inmunoaglutinacion para detectar la enfermedad de Chagas, mediante un nuevo complejo de latex – (antigeno quimerico recombinante, muestra una mayor discriminacion entre las respuestas al suero control positivo y al suero control negativo. Metodos Se determinaron las siguientes variables: (i) el mecanismo de sensibilizacion, (ii) el agente emulsificador utilizado para la desabsorcion de proteinas, (iii) el tiempo de reaccion, (iv) la fuerza ionica del medio de reaccion, (v) la concentracion de particulas, (vi) la presencia de agentes bloqueantes (vii) la presencia de polietilenglicol como potenciador de la reaccion, y (viii) las concentraciones de antigeno y anticuerpo. La busqueda de condiciones optimas se realizo cambiando una variable a la vez. Con este objetivo, las particulas monodispersas de latex, sensibilizadas con una proteina quimerica recombinante (CP1) fueron expuestas a diferentes condiciones. Se realizo un seguimiento a la reaccion de aglutinacion, midiendo los cambios en absorbancia optica mediante turbidimetria. Resultados La discriminacion maxima entre sueros negativos y positivos se obtuvo en un tiempo de reaccion de 5 minutos cuando complejos de latex, con una concentracion de 2.90 mg/m2 de proteina asociada covalentemente, se pusieron en contacto con suero sin diluir en una solucion tampon de borato pH 8–20 mm con glicina (0.1 m) y polietileneglicol 8000 (3% w/v). Finalmente, el complejo latex-proteina se probo bajo las condiciones optimas, con un panel de sueros T. cruzi-positivos, sueros positivos para leishmaniosis y sueros negativos para ambos parasitos. Conclusion La prueba de inmunoaglutinacion basada en el complejo latex-CP1 podria utilizarse como un metodo de deteccion de la enfermedad de Chagas. Esta prueba es rapida, facil de implementar y podria utilizarse bajo condiciones de campo; pero sus resultados deberian confirmarse mediante tecnicas de referencia tales como ELISA, HAI e IFI.

This paper presents an optical method for real-time monitoring of protein adsorption using porous silicon self-supported microcavities as a label-free detection platform. The study combines an experimental approach with a physical model for the adsorption process. The proposed model agrees well with experimental observations, and provides information about the kinetics of diffusion and adsorption of proteins within the pores, which will be useful for future experimental designs Fil: Lasave, Liliana Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Urteaga, Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina. Universidad Nacional del Litoral; Argentina Fil: Koropecki, Roberto Roman. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina. Universidad Nacional del Litoral; Argentina Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Arce, Roberto Delio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina

The physical adsorption and the chemical coupling of recombinant proteins of Trypanosoma cruzi onto polystyrene and core-shell carboxylated particles were respectively investigated with the ultimate aim of producing latex-protein complexes to be used in an immunoagglutination assay able to detect the Chagas disease. To this effect, two single proteins (RP1 and RP5) and a multiepitope protein derived from three antigenic peptides (CP2) were evaluated, and sensitizations were carried out at different pHs. The maximum physical adsorption was produced at pHs close to the protein isoelectric point (i.e., pH 6 for RP5 and pH 5 for RP1 and CP2). High fractions of antigens were chemically bound to the carboxyl groups, and the highest surface density of linked protein was also observed at pHs close to the protein isoelectric point. The three latex-protein complexes obtained by covalent coupling at such pHs were tested with sera from a panel of 16 infected and 16 non-infected patients. In the immunoagglutination assays, the latex-CP2 complex produced the best discrimination between positive and negative sera. Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina Fil: Caudana, Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina Fil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina Fil: Marcipar, Ivan Sergio. Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentina

The acute phase recombinant protein of Toxoplasma gondii P22Ag was expressed and purified and thehomogenate of the parasite was obtained from an infected mouse. These antigens were used to producelatex-protein complexes (LPC) through physical adsorption and chemical coupling onto different latexes,with the aim of producing immunoagglutination (IA) reagents able to detect recently acquiredtoxoplasmosis. Polystyrene and ?core-shell? latexes where employed, exhibiting varied particle size,functionality (carboxyl or epoxy), and charge density. In sensitization experiments for producing LPC, therecombinant protein showed better coupling efficiency onto the particles surface than the homogenateand this could be explained by the complex mixture of the homogenate, which includes a large numberof proteins of different molecular mass, isoelectric points, and hydrophobicity. The synthesized LPC wereemployed in IA assays. To this effect, the agglutination reaction was followed by measuring the changesin the optical absorbance by turbidimetry. Experiments against control sera were performed to evaluatethe performance of various LPC and it was observed that the IA test based on P22Ag and thecarboxylated latex of 350 nm of particle diameter allowed a good discrimination between acute sera andchronic/negative ones. The proposed test is cheap, rapid, and easy to implement. Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Costa, Juan Gabriel. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

Background: P35 and P22 Toxoplasma gondii proteins are recognized by specific IgG at the early infection stage, making them ideal for acute toxoplasmosis pregnancy control. Both proteins have been studied to discriminate between acute and chronic toxoplasmosis. However, results were hardly comparable because different protein obtainment procedures led to different antigens, the referencepanels used were not optimally typified, and avidity tests were either not performed or narrowly examined. Methods: We bioinformatically predicted P35 andP22 regions with the highest density of epitopes, and expressed them in pET32/BL21DE3 alternative expression system, obtaining the soluble proteins rP35a and rP22a. We assessed their diagnostic performance using pregnant woman serum samples typified as: not infected, NI (IgG−, IgM−), typical-chronic, TC (IgM−, IgG+), presumably acute, A (IgG+, IgM+, low-avidity IgG), and recentlychronic, RC (IgG+, IgM+, high-avidity IgG). Fil: Costa, Juan Gabriel. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Peverengo, Luz. Universidad Nacional del Litoral; Argentina Fil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Dalla Fontana, María. Laboratorio Central de la Provincia de Santa Fe; Argentina Fil: Lagier, Claudia Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina Fil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The particle size distribution (PSD) of dilute latex was estimated through a general regression neural network (GRNN) that was supplied with PSD average diameters derived from multiangle dynamic light scattering (MDLS) measurements. The GRNN was trained with a large set of measurements that were simulated from unimodal normal-logarithmic distributions representing the PSDs of polystyrene (PS) latexes. The proposed method was first tested through three simulated examples involving different PSD shapes, widths, and diameter ranges. Then the GRNN was employed to estimate the PSD of two PS samples; a latex standard of narrow PSD and known nominal diameter, and a latex synthesized in our laboratory. Both samples were also characterized through independent techniques (capillary hydrodynamic fractionation, transmission electron microscopy, and disc centrifugation). For comparison, all examples were solved by numerical inversion of MDLS measurements through a Tikhonov regularization technique. The PSDs estimated by the GRNN gave more accurate results than those obtained through other conventional techniques. The proposed method is a simple, effective, and robust tool for characterizing unimodal PSDs. Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Stegmayer, Georgina. Universidad Tecnologica Nacional. Facultad Regional Santa Fe. Centro de Investigacion y Desarrollo de Ingenieria en Sistemas de Informacion; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Clementi, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Leiza, José. Universidad del País Vasco; España Fil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina. Universidad Tecnologica Nacional. Facultad Regional Santa Fe. Centro de Investigacion y Desarrollo de Ingenieria en Sistemas de Informacion; Argentina

A novel immunodiagnosis reagent for detecting the Chagas Disease was developed, by chemical coupling of antigen Ag36 of Trypanosoma cruzi onto two (carboxylated and core-shell) latexes. The coupling reactions involved the use of a carbodiimide intermediate. Bovine serum albumin (BSA) was used as a model protein for determining the appropriate conditions for its physical and chemical coupling. BSA showed an increased adsorption onto the base carboxylated latexes, with respect to a PS latex without carboxyl groups. The chemical bonding experiments only involved the carboxylated latexes. With BSA, the final density of covalently bound protein was 2.30 mg/m(2). In addition, around 55% of the total linked protein was chemically coupled, and the reaction was little affected by the pH. With Ag36, the final density of covalently bound protein was 2.44 mg/m(2), around 80% of the total linked protein was chemically coupled, and the chemical coupling was maximum at pH = 5 (i.e., close to the isoelectric point).

The grafting of casein to acrylic polymers is needed to fully exploit the possibilities of hybrid nanoparticles containing such materials. The explored alternative up to now, based on the primary radical formation onto the protein chains through the redox initiation between the casein amino groups and a hydroperoxide, produces a limited degree of compatibilization to guarantee the synergic effect between both components. A novel strategy that overcomes these limitations is presented. The strategy is based on the use of a crosslinkable casein, which in addition to the characteristic grafting capacity by redox initiation with hydroperoxides, contains acrylic groups able to radically polymerize. The degree of grafting of both acrylic and casein is significantly increased by functionalizing casein with varied amounts of pendant propagating points per protein. Moreover, it is observed that the improved degree of compatibility obtained when using the crosslinkable casein considerably enhanced the properties of the acrylic/casein films, such as their mechanical behavior and resistance to both water and organic solvent. Fil: Picchio, Matías Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Barandiaran, María J.. Universidad del País Vasco; España Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

Multiangle dynamic light scattering (MDLS) and turbidimetry (T) were applied (both individually and combined) for determining the contamination by larger particles of two almost-uniform polystyrene (PS) latices. Latex 1 was synthesized in our laboratories, and it contained a main population diameter of 340 nm together with a small fraction of larger particles. This latex was used as the base material for producing an immunoassay kit. Latex 2 was obtained by a simple blend of two uniform PS standards. The proposed data treatment calculates the diameter and number fraction of the large particles contamination assuming that the PSDs are bimodal. The calculation involves minimizing the errors between the measurements and their theoretical predictions. When analyzed by combined MDLS–T, the contamination of Latex 1 involved number fraction 0.6% and particle diameter 865 nm. The T average diameter is a function of the measurement wavelength, and the highest deviations of this average to an increasing contamination by large particles were always observed at the higher wavelengths. The DLS average diameter is a function of the measurement angle, but in this case it is impossible to determine a priori the angle of observation that provides the largest deviation of this average diameter to an increasing contamination. Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Meira, Gregorio Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

Multi-angle dynamic light scattering (DLS) provides a better estimate of the particle size distribution (PSD) than single-angle DLS. However, multi-angle data treatment requires to appropriately weight each autocorrelation measurement prior to calculating the PSD. The weighting coefficients may be directly obtained from: i) the autocorrelation baselines, or ii) independent measurement of the average light intensity by elastic light scattering. However, the propagation of errors associated to such procedures may intolerably corrupt the PSD estimate. In this work, an alternative recursive least squares calculation is proposed that estimates the weighting coefficients on the basis of the complete autocorrelation measurement. The method was validated through a numerical example that simulates the analysis of a polystyrene latex with a bimodal PSD; and with “measurements” taken at 10 detection angles. The ill-conditioned nature of the problem determines that the “true” PSD cannot be recuperated, even in the absence of errors. A sensitivity analysis was carried out to determine the effect of errors in the weighting coefficients on the PSD recuperations. Fil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: González, Verónica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Meira, Gregorio Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

We have evaluated blood samples of chronic and congenital Trypanosoma cruzi-infected patients from the city of Reconquista located in the northeast of Argentina where no information was previously obtained about the genotype of infecting parasites. Fourteen samples of congenital and 19 chronical patients were analyzed by hybridization with DNA probes of minicircle hypervariable regions (mHVR). In congenital patients, 50% had single infections with TcIId, 7% single infections with TcIIe, 29% mixed infections with TcIId/e, and 7% had mixed infections with TcIId/b and 7% TcIId/b, respectively. In Chronical patients, 52% had single infections with TcIId, 11% single infections with TcIIe, 26% had mixed infections with TcIId/e, and 11% had non-identified genotypes. With these samples, we evaluated the minicircle lineage-specific polymerase chain reaction assay (MLS-PCR), which involves a nested PCR to HVR minicircle sequences and we found a correlation with hybridization probes of 96.4% for TcIId and 54.8% for TcIIe.

This article describes the physical adsorption and the chemical coupling of 3 antigenic proteins of Trypanosoma cruzi onto polystyrene (PS) based latexes to be used as novel immunodiagnosis reagents for detecting the Chagas disease. The coupled proteins were a homogenate of T. cruzi, or a recombinant protein (either Ag36 or CP1). With the homogenate, between 30 and 60% of the total-linked protein was chemically coupled, showing a small dependence with the pH. For Ag36 and CP1, around 90% of the total-linked protein was chemically coupled, with a maximum coupling at pH 5 (i.e., close to the isoelectric point). The chemical coupling of CP1 was less affected by the pH than the coupling of Ag36.

The aim of this work was to determine the best strategy to display antigens (Ags) on immunochemical devices to improve test selectivity and sensitivity. We comparatively evaluated five Trypanosoma cruzi antigenic recombinant peptides, chose the three more sensitive ones, built up chimeras bearing these selected Ags, and systematically compared by enzyme-linked immunosorbent assay the performance of the assortments of those peptides with that of the multiepitope constructions bearing all those peptides lineally fused. The better-performing Ags that were compared included peptides homologous to the previously described T. cruzi flagellar repetitive Ag (here named RP1), shed acute-phase Ag (RP2), B13 (RP5), and the chimeric recombinant proteins CP1 and CP2, bearing repetitions of RP1-RP2 and RP1-RP2-RP5, respectively. The diagnostic performances of these Ags were assessed for discrimination efficiency by the formula +OD/cutoff value (where +OD is the mean optical density value of the positive serum samples tested), in comparison with each other either alone, in mixtures, or as peptide-fused chimeras and with total parasite homogenate (TPH). The discrimination efficiency values obtained for CP1 and CP2 were 25% and 52% higher, respectively, than those of their individual-Ag mixtures. CP2 was the only Ag that showed enhanced discrimination efficiency between Chagas´ disease-positive and -negative samples, compared with TPH. This study highlights the convenience of performing immunochemical assays using hybrid, single-molecule, chimeric Ags instead of peptide mixtures. CP2 preliminary tests rendered 98.6% sensitivity when evaluated with a 141-Chagas´ disease-positive serum sample panel and 99.4% specificity when assessed with a 164-Chagas´ disease-negative serum sample panel containing 15 samples from individuals infected with Leishmania spp. Fil: Camussone, Cecilia María. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: González, Verónica Doris Guadalupe. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Belluzo, María Soledad. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Pujato, Nazarena. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ribone, María Élida. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química Analítica; Argentina Fil: Lagier, Claudia Marina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química Analítica; Argentina Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

This article investigates the synthesis of two (monodisperse, carboxylated, and core-shell) latexes, through a batch and a semibatch emulsion copolymerizations of styrene (St) and methacrylic acid (MAA) onto polystyrene latex seeds. A mathematical model of the process was developed that predicts conversion, average particle size, and surface density of carboxyl groups. The model was adjusted to the batch reaction measurements, and then it was used in the design of the semibatch experiment. The semibatch reaction involved an initial homopolymerization of St followed by instantaneous addition of MAA-St-initiator. Compared with the batch reaction results, the semibatch policy more than doubled the surface density of carboxyl groups. The second part of this series describes the development of an immunodiagnosis latex-protein complex for detecting the Chagas disease, by coupling an antigen of Trypanosoma cruzi onto the produced carboxylated latexes.