Search

Your search keyword '"Jain, A. (Ashok)"' showing total 21 results
Start Over Author "Jain, A. (Ashok)"
21 results on '"Jain, A. (Ashok)"'

3. The development of biologics for use in translational medicine

Author

Jain, Saurabh Ashok, Hupp, Ted, and Argyle, David

Subjects
572, synuclein, CD20
Abstract

In this research, we developed a wide range of biological tools against two distinct targets from future diagnostic or therapeutic points of view. Firstly, we demonstrate that sporadic canine B cell lymphoma mimics the features of human equivalents which in turn will be advantageous for development of canine as well as human therapeutics. With a comparative oncological approach, here we developed a monoclonal antibody (NCD1.2) against canine CD20 which also binds to its human counterpart. Using flow cytometry and tissue microarray, we show that NCD1.2 binds specifically to canine B cell lymphomas (CD20+) and not T-cell lymphoma (CD20-). We also cloned scFv scaffold by linking variable heavy and light chains from NCD1.2 hybridoma by a serine-glycine linker to see if it was active as a biological tool for future therapeutics. Intriguingly, we obtained two different kappa light chains from a single hybridoma cell (scFv3 and scFv7) after antibody phage display. These scFvs were cloned into mammalian vectors for expression in CHO cells and ADEPT - CpG2 vector for yeast expression to see if the activity of these scFvs was retained. Our data suggests that recombinant anti-CD20 scFv might be a useful tool for bioconjugate directed immunotherapies in comparative medicine. Secondly, in addition to mAbs we also developed peptide aptamers which are seldom described but have become attractive agents that typically target a specific biomolecule of interest. Parkinson’s disease (PD) is characterized by formation of lewy bodies (inclusion bodies) in the substantia nigra and the major content of lewy bodies is α-synuclein. To begin with we made recombinant α-synuclein and biophysically characterize this protein under different conditions on a native gel. We also performed Circular dichroism to look at its structure and demonstrate that α helicity could be achieved in presence of SDS. The aim of this project was to develop peptide aptamers, mAbs to α-synuclein, map the binding sites onto the peptides derived from the protein and also on recombinant protein. Further we demonstrated the development of biological tools and their potential ability against α-synuclein in α-synuclein expressing cell lines from future PD therapeutic perspective. Monoclonal antibodies were developed and mAb (3.1) was found to be immunopositive for α-synuclein in parts of kidney and brain. Moreover to estimate the oligomeric state of α-synuclein, we developed assays such as co-transfection of two different constructs i.e. cherry and GFP tagged α-synuclein and Proximity ligation assay to show its self - interaction. Peptide phage display screening (NEB Ph.D. 12 mer library) on recombinant WT α-synuclein was performed to identify aptamers and ultimately novel binding proteins. The peptides were selected based on iteration number and out of the selected panel of peptides; SHACWWDECTGS was found to effectively bind α-synuclein using ELISA. Scanning of peptide GDGNSVLKPGNW (highest iteration number) led to identification of interacting proteins with α-synuclein. Thus in conclusion, we show the validation of different antibody scaffolds and peptide aptamers which could be useful tools from future therapeutics point of view against two well characterized antigens in B cell lymphoma and Parkinson’s disease, respectively.

Published
2015

5. Discovery and functional prioritization of Parkinson's disease candidate genes from large-scale whole exome sequencing

Author

Jansen, I. (Iris), Ye, H. (Hui), Heetveld, S. (Sasja), Lechler, M.C. (Marie C.), Michels, H. (Helen), Seinstra, R.I. (Renée I.), Lubbe, S.J. (Steven J.), Drouet, V. (Valérie), Lesage, S. (Suzanne), Majounie, E. (Elisa), Gibbs, J.R. (J.Raphael), Nalls, M.A. (Michael), Ryten, M. (Mina), Botia, J.A. (Juan A.), Vandrovcova, J. (Jana), Simón-Sánchez, J. (Javier), Castillo-Lizardo, M. (Melissa), Rizzu, P. (Patrizia), Blauwendraat, C. (Cornelis), Chouhan, A.K. (Amit K.), Li, Y. (Yarong), Yogi, P. (Puja), Amin, N. (Najaf), Duijn, C.M. (Cornelia) van, Morris, H.R. (Huw R.), Brice, A. (Alexis), Singleton, A. (Andrew), David, D.C. (Della C.), Nollen, E.A. (Ellen A.), Jain, A. (Ashok), Shulman, J.M., Heutink, P. (Peter), Hernandez, D.G. (Dena), Arepalli, S. (Sampath), Brooks, J. (Janet), Price, R. (Ryan), Nicolas, A. (Aude), Chong, S. (Sean), Cookson, M.R. (Mark), Dillman, A. (Allissa), Moore, M. (Matt), Traynor, B.J. (Bryan), Plagnol, V. (Vincent), Nicholas W Wood, Sheerin, U.-M. (Una-Marie), Jose M Bras, Charlesworth, K. (Kate), Gardner, M. (Mac), Guerreiro, R. (Rita), Trabzuni, D. (Danyah), Hardy, J. (John), Sharma, M., Saad, M. (Mohamad), Javier Simón-Sánchez, Schulte, C. (Claudia), Corvol, J.C. (Jean-Christophe), Dürr, A. (Alexandra), Vidailhet, M. (M.), Sveinbjörnsdóttir, S. (Sigurlaug), Barker, R.A. (Roger), Caroline H Williams-Gray, Ben-Shlomo, Y., Berendse, H.W. (Henk W.), Dijk, K.D. (Karin) van, Berg, D. (Daniela), Brockmann, K., Wurster, K.D. (Kathrin), Mätzler, W. (Walter), Gasser, T. (Thomas), Martinez, M. (Maria), Bie, R.M.A. (Rob) de, Biffi, A. (Alessandro), Velseboer, D. (Daan), Bloem, B.R. (Bastiaan), Post, B. (Bart), Wickremaratchi, M. (Mirdhu), Warrenburg, B. (Bart) van de, Bochdanovits, Z. (Zoltan), Bonin, M. (Malte) von, Pétursson, H. (Hjörvar), Riess, O. (Olaf), Burn, D.J. (David), Lubbe, S. (Steven), Cooper, J.M. (J Mark), McNeill, N.H. (Nathan), Schapira, A. (Anthony), Lungu, C. (Codrin), Chen, H. (Honglei), Dong, J. (Jing), Chinnery, P.F. (Patrick F.), Hudson, G. (Gavin), Clarke, C.E. (Carl E.), Moorby, C. (Catriona), Counsell, C. (Carl), Damier, P. (Philippe), Dartigues, J.-F., Deloukas, P. (Panagiotis), Gray, E. (Emma), Edkins, T. (Ted), Hunt, S.E. (Sarah E.), Potter, S.C. (Simon), Tashakkori-Ghanbaria, A. (Avazeh), Deuschl, G. (Günther), Lorenz, D. (Delia), Dexter, D.T. (David), Durif, F. (Frank), Evans, J. (Jonathan Mark), Langford, C. (Cordelia), Foltynie, T. (Thomas), Goate, A.M. (Alison), Harris, C. (Clare), Hilten, J.J. (Jacobus) van, Hofman, A. (Albert), Hollenbeck, J.R. (John R.), Holton, J.L. (Janice), Hu, M. (Michele), Huang, X. (Xiaohong), Illig, T. (Thomas), Jónsson, P.V. (Pálmi), Lambert, J.-C., O'Sullivan, S.S. (Sean), Revesz, T. (Tamas), Shaw, K. (Karen), Lees, A.J. (Andrew), Lichtner, P. (Peter), Limousin, P. (Patricia), Lopez, G., Escott-Price, V. (Valentina), Pearson, J. (Justin), Williams, N. (Nigel), Mudanohwo, E. (Ese), Perlmutter, J.S. (Joel), Pollak, P. (Pierre), Rivadeneira Ramirez, F. (Fernando), Uitterlinden, A.G. (André), Sawcer, S.J. (Stephen), Scheffer, H. (Hans), Shoulson, I. (Ira), Shulman, L. (Lee), Smith, C. (Colin), Walker, R. (Robert), Spencer, C.C.A. (Chris C.), Strange, A. (Amy), Stefansson, H. (Hreinn), Bettella, F. (Francesco), Zwart, J-A. (John-Anker), Stockton, J.D. (Joanna D.), Talbot, D., Tanner, C.M. (Carlie), Tison, F. (François), Winder-Rhodes, S. (Sophie), Bhatia, K.P. (Kailash), Jansen, I. (Iris), Ye, H. (Hui), Heetveld, S. (Sasja), Lechler, M.C. (Marie C.), Michels, H. (Helen), Seinstra, R.I. (Renée I.), Lubbe, S.J. (Steven J.), Drouet, V. (Valérie), Lesage, S. (Suzanne), Majounie, E. (Elisa), Gibbs, J.R. (J.Raphael), Nalls, M.A. (Michael), Ryten, M. (Mina), Botia, J.A. (Juan A.), Vandrovcova, J. (Jana), Simón-Sánchez, J. (Javier), Castillo-Lizardo, M. (Melissa), Rizzu, P. (Patrizia), Blauwendraat, C. (Cornelis), Chouhan, A.K. (Amit K.), Li, Y. (Yarong), Yogi, P. (Puja), Amin, N. (Najaf), Duijn, C.M. (Cornelia) van, Morris, H.R. (Huw R.), Brice, A. (Alexis), Singleton, A. (Andrew), David, D.C. (Della C.), Nollen, E.A. (Ellen A.), Jain, A. (Ashok), Shulman, J.M., Heutink, P. (Peter), Hernandez, D.G. (Dena), Arepalli, S. (Sampath), Brooks, J. (Janet), Price, R. (Ryan), Nicolas, A. (Aude), Chong, S. (Sean), Cookson, M.R. (Mark), Dillman, A. (Allissa), Moore, M. (Matt), Traynor, B.J. (Bryan), Plagnol, V. (Vincent), Nicholas W Wood, Sheerin, U.-M. (Una-Marie), Jose M Bras, Charlesworth, K. (Kate), Gardner, M. (Mac), Guerreiro, R. (Rita), Trabzuni, D. (Danyah), Hardy, J. (John), Sharma, M., Saad, M. (Mohamad), Javier Simón-Sánchez, Schulte, C. (Claudia), Corvol, J.C. (Jean-Christophe), Dürr, A. (Alexandra), Vidailhet, M. (M.), Sveinbjörnsdóttir, S. (Sigurlaug), Barker, R.A. (Roger), Caroline H Williams-Gray, Ben-Shlomo, Y., Berendse, H.W. (Henk W.), Dijk, K.D. (Karin) van, Berg, D. (Daniela), Brockmann, K., Wurster, K.D. (Kathrin), Mätzler, W. (Walter), Gasser, T. (Thomas), Martinez, M. (Maria), Bie, R.M.A. (Rob) de, Biffi, A. (Alessandro), Velseboer, D. (Daan), Bloem, B.R. (Bastiaan), Post, B. (Bart), Wickremaratchi, M. (Mirdhu), Warrenburg, B. (Bart) van de, Bochdanovits, Z. (Zoltan), Bonin, M. (Malte) von, Pétursson, H. (Hjörvar), Riess, O. (Olaf), Burn, D.J. (David), Lubbe, S. (Steven), Cooper, J.M. (J Mark), McNeill, N.H. (Nathan), Schapira, A. (Anthony), Lungu, C. (Codrin), Chen, H. (Honglei), Dong, J. (Jing), Chinnery, P.F. (Patrick F.), Hudson, G. (Gavin), Clarke, C.E. (Carl E.), Moorby, C. (Catriona), Counsell, C. (Carl), Damier, P. (Philippe), Dartigues, J.-F., Deloukas, P. (Panagiotis), Gray, E. (Emma), Edkins, T. (Ted), Hunt, S.E. (Sarah E.), Potter, S.C. (Simon), Tashakkori-Ghanbaria, A. (Avazeh), Deuschl, G. (Günther), Lorenz, D. (Delia), Dexter, D.T. (David), Durif, F. (Frank), Evans, J. (Jonathan Mark), Langford, C. (Cordelia), Foltynie, T. (Thomas), Goate, A.M. (Alison), Harris, C. (Clare), Hilten, J.J. (Jacobus) van, Hofman, A. (Albert), Hollenbeck, J.R. (John R.), Holton, J.L. (Janice), Hu, M. (Michele), Huang, X. (Xiaohong), Illig, T. (Thomas), Jónsson, P.V. (Pálmi), Lambert, J.-C., O'Sullivan, S.S. (Sean), Revesz, T. (Tamas), Shaw, K. (Karen), Lees, A.J. (Andrew), Lichtner, P. (Peter), Limousin, P. (Patricia), Lopez, G., Escott-Price, V. (Valentina), Pearson, J. (Justin), Williams, N. (Nigel), Mudanohwo, E. (Ese), Perlmutter, J.S. (Joel), Pollak, P. (Pierre), Rivadeneira Ramirez, F. (Fernando), Uitterlinden, A.G. (André), Sawcer, S.J. (Stephen), Scheffer, H. (Hans), Shoulson, I. (Ira), Shulman, L. (Lee), Smith, C. (Colin), Walker, R. (Robert), Spencer, C.C.A. (Chris C.), Strange, A. (Amy), Stefansson, H. (Hreinn), Bettella, F. (Francesco), Zwart, J-A. (John-Anker), Stockton, J.D. (Joanna D.), Talbot, D., Tanner, C.M. (Carlie), Tison, F. (François), Winder-Rhodes, S. (Sophie), and Bhatia, K.P. (Kailash)

Abstract

Background: Whole-exome sequencing (WES) has been successful in identifying genes that cause familial Parkinson's disease (PD). However, until now this approach has not been deployed to study large cohorts of unrelated participants. To discover rare PD susceptibility variants, we performed WES in 1148 unrelated cases and 503 control participants. Candidate genes were subsequently validated for functions relevant to PD based on parallel RNA-interference (RNAi) screens in human cell culture and Drosophila and C. elegans models. Results: Assuming autosomal recessive inheritance, we identify 27 genes that have homozygous or compound heterozygous loss-of-function variants in PD cases. Definitive replication and confirmation of these findings were hindered by potential heterogeneity and by the rarity of the implicated alleles. We therefore looked for potential genetic interactions with established PD mechanisms. Following RNAi-mediated knockdown, 15 of the genes modulated mitochondrial dynamics in human neuronal cultures and four candidates enhanced α-synuclein-induced neurodegeneration in Drosophila. Based on complementary analyses in independent human datasets, five functionally validated genes-GPATCH2L, UHRF1BP1L, PTPRH, ARSB, and VPS13C-also showed evidence consistent with genetic replication. Conclusions: By integrating human genetic and functional evidence, we identify several PD susceptibility gene candidates for further investigation. Our approach highlights a powerful experimental strategy with broad applicability for future studies of disorders with complex genetic etiologies.

Published
2017
Full Text
View/download PDF

6. Development of biologics for use in translational medicine

Author

Jain, Saurabh Ashok, Hupp, Ted, Argyle, David, and other

Subjects
synuclein, CD20
Abstract

In this research, we developed a wide range of biological tools against two distinct targets from future diagnostic or therapeutic points of view. Firstly, we demonstrate that sporadic canine B cell lymphoma mimics the features of human equivalents which in turn will be advantageous for development of canine as well as human therapeutics. With a comparative oncological approach, here we developed a monoclonal antibody (NCD1.2) against canine CD20 which also binds to its human counterpart. Using flow cytometry and tissue microarray, we show that NCD1.2 binds specifically to canine B cell lymphomas (CD20+) and not T-cell lymphoma (CD20-). We also cloned scFv scaffold by linking variable heavy and light chains from NCD1.2 hybridoma by a serine-glycine linker to see if it was active as a biological tool for future therapeutics. Intriguingly, we obtained two different kappa light chains from a single hybridoma cell (scFv3 and scFv7) after antibody phage display. These scFvs were cloned into mammalian vectors for expression in CHO cells and ADEPT - CpG2 vector for yeast expression to see if the activity of these scFvs was retained. Our data suggests that recombinant anti-CD20 scFv might be a useful tool for bioconjugate directed immunotherapies in comparative medicine. Secondly, in addition to mAbs we also developed peptide aptamers which are seldom described but have become attractive agents that typically target a specific biomolecule of interest. Parkinson’s disease (PD) is characterized by formation of lewy bodies (inclusion bodies) in the substantia nigra and the major content of lewy bodies is α-synuclein. To begin with we made recombinant α-synuclein and biophysically characterize this protein under different conditions on a native gel. We also performed Circular dichroism to look at its structure and demonstrate that α helicity could be achieved in presence of SDS. The aim of this project was to develop peptide aptamers, mAbs to α-synuclein, map the binding sites onto the peptides derived from the protein and also on recombinant protein. Further we demonstrated the development of biological tools and their potential ability against α-synuclein in α-synuclein expressing cell lines from future PD therapeutic perspective. Monoclonal antibodies were developed and mAb (3.1) was found to be immunopositive for α-synuclein in parts of kidney and brain. Moreover to estimate the oligomeric state of α-synuclein, we developed assays such as co-transfection of two different constructs i.e. cherry and GFP tagged α-synuclein and Proximity ligation assay to show its self - interaction. Peptide phage display screening (NEB Ph.D. 12 mer library) on recombinant WT α-synuclein was performed to identify aptamers and ultimately novel binding proteins. The peptides were selected based on iteration number and out of the selected panel of peptides; SHACWWDECTGS was found to effectively bind α-synuclein using ELISA. Scanning of peptide GDGNSVLKPGNW (highest iteration number) led to identification of interacting proteins with α-synuclein. Thus in conclusion, we show the validation of different antibody scaffolds and peptide aptamers which could be useful tools from future therapeutics point of view against two well characterized antigens in B cell lymphoma and Parkinson’s disease, respectively.

Published
2015

15. Antibody titers in Group O platelet donors.

Author

Tendulkar, Anita Amar, Jain, Puneet Ashok, and Velaye, Sanjay

Subjects
*ABO blood group system, *BLOOD platelets, *CHEMICAL reagents, *DIAGNOSTIC errors, *IMMUNOGLOBULINS, *LONGITUDINAL method, *POPULATION, *STATISTICAL sampling, *STATISTICS, *DATA analysis, *DESCRIPTIVE statistics
Abstract

Background and Objectives: The occurrence of hemolysis due to transfusion of ABO plasma-incompatible platelets (PLTs) is challenging. There has been no consensus for critical antibody titers in the transfusion community. This study was conducted to understand the trends of anti-A and anti-B antibody titer levels in O group donors and to identify any specific patterns of distribution in relation to age and gender. Materials and Methods: A total of 1635 Group O PLT donors were randomly selected for this prospective study. Serial 2-fold doubling dilutions were prepared for each sample to calculate the titer of anti-A and anti-B in a standard 96 well micro-plate. Tube technique was used for comparison with the microplate method for 100 samples. Results: Out of 1635 donors, 1430 (87.46%) were males and 205 (12.54%) were females. The median titer for anti-A and anti-B was 128 with range from 4 to 2048. Spearman's correlation coefficient for microplate versus tube technique was estimated to be 0.803 (P < 0.01, two-tailed). 57.12% and 51.19% of all donors had titers ≥128 for anti-A and anti-B, respectively. The geometric mean of anti-A and anti-B was 155.7 and 137.28, respectively. The titers were significantly higher (P < 0.001) in female donors. An inverse relation between titer levels and age was seen. Conclusion: Microplate can be used to perform titers in resource-constrained settings. Screening for critical titers in O group donors is essential as they are more implicated in hemolytic transfusion reactions. In the absence of a global consensus on this topic, institutes may need to formulate their own guidelines on handling ABO plasma-incompatible PLT transfusions. [ABSTRACT FROM AUTHOR]

Published
2017
Full Text
View/download PDF

16. Suppression of HBV by Tenofovir in HBV/HIV Coinfected Patients: A Systematic Review and Meta-Analysis

Author

Price, H. (Huw), Dunn, D. (David), Pillay, D. (Deenan), Bani-Sadr, F. (Firouze), Vriessluijs, T.E.M.S. (Theodora) de, Jain, A. (Ashok), Kuzushita, N. (Noriyoshi), Mauss, S. (Stefan), Núñez, M.J. (Marina), Nüesch, E. (Eveline), Peters, M.G. (Marion), Reiberger, T. (Thomas), Stephan, C. (Carsten), Tan, L. (Lionel), Gilson, E. (Eric), Price, H. (Huw), Dunn, D. (David), Pillay, D. (Deenan), Bani-Sadr, F. (Firouze), Vriessluijs, T.E.M.S. (Theodora) de, Jain, A. (Ashok), Kuzushita, N. (Noriyoshi), Mauss, S. (Stefan), Núñez, M.J. (Marina), Nüesch, E. (Eveline), Peters, M.G. (Marion), Reiberger, T. (Thomas), Stephan, C. (Carsten), Tan, L. (Lionel), and Gilson, E. (Eric)

Abstract

Background:Hepatitis B coinfection is common in HIV-positive individuals and as antiretroviral therapy has made death due to AIDS less common, hepatitis has become increasingly important. Several drugs are available to treat hepatitis B. The most potent and the one with the lowest risk of resistance appears to be tenofovir (TDF). However there are several questions that remain unanswered regarding the use of TDF, including the proportion of patients that achieves suppression of HBV viral load and over what time, whether suppression is durable and whether prior treatment with other HBV-active drugs such as lamivudine, compromises the efficacy of TDF due to possible selection of resistant HBV strains.Methods:A systematic review and meta-analysis following PRISMA guidelines and using multilevel mixed effects logistic regression, stratified by prior and/or concomitant use of lamivudine and/or emtricitabine.Results:Data was available from 23 studies including 550 HBV/HIV coinfected patients treated with TDF. Follow up was for up to seven years but to ensure sufficient power the data analyses were limited to three years. The overall proportion achieving suppression of HBV replication was 57.4%, 79.0% and 85.6% at one, two and three years, respectively. No effect of prior or concomitant 3TC/FTC was shown. Virological rebound on TDF treatment was rare.Interpretation:TDF suppresses HBV to undetectable levels in the majority of HBV/HIV coinfected patients with the proportion fully suppressed continuing to increase during continuous treatment. Prior treatment with 3TC/FTC does not compromise efficacy of TDF treatment. The use of combination treatment with 3TC/FTC offers no significant benefit over TDF alone.

Published
2013
Full Text
View/download PDF

17. Secondary peritonitis caused by Streptomyces viridis

Author

Datta, P. (Priya), Arora, S. (Shilpa), Jain, A. (Ashok), Chander, J. (Jagdish), Sande, W.W.J. (Wendy) van de, Datta, P. (Priya), Arora, S. (Shilpa), Jain, A. (Ashok), Chander, J. (Jagdish), and Sande, W.W.J. (Wendy) van de

Abstract

Streptomyces organisms are soil inhabitants rarely causing nonmycetomic infections. We describe a case of secondary peritonitis caused by Streptomyces viridis in a chronic alcoholic patient who presented

Published
2012
Full Text
View/download PDF

21. Design and LENS® Fabrication of Bi-metallic Cu-H13 Tooling for Die Casting

Author

Jain, Akshay Ashok

Subjects
Engineering, Industrial Engineering, Mechanical Engineering, Metallurgy, Die Casting, Bi-metallic tooling, Copper, H13, Die Casting Tooling, Ansys, Finite Element Analysis, FEA, Thermal Fatigue Die Casting, Laser Engineered Net Shaping, Laser Additive Manufacturing, Copper-H13, H13-Copper, Cu-H13, H13-Cu, Dunk Test
Abstract

Thermal fatigue is one of the most common causes leading to die failure in die casting. This thesis investigates and presents the results of thermal fatigue life in a bi-metallic H13-Copper die which can be manufactured using laser additive manufacturing technologies now commercially available. Using finite element method, computational models are developed to simulate the thermal fatigue tests of Wallace (Benedyk, Moracz, and Wallace 1970). Numerical solutions to the thermal-mechanical problem are obtained. The solutions include temperature, strain and stress distributions within the test sample. Solutions were obtained for varying amounts of copper in the test sample geometry. Results from the pure H13 sample computational model compared very well with experimental values obtained by Wallace (Benedyk, Moracz, and Wallace 1970).The maximum temperature reached by the test sample is shown to decrease with increasing amounts of copper. The fatigue life is calculated using the `method of universal slopes’ which relates the calculated cyclic strain ranges to the number of cycles necessary for fatigue crack initiation. The specimen geometry consisting of a half thickness of Cu and the other half thickness of H13 at the thinnest point in the full cross-section of the wall thickness was shown to provide the best balance between thermal and fatigue life performance.

Published
2013

Catalog

Books, media, physical & digital resources
See catalog results