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Magnetic nanoparticles are an attractive bioseparation tool due to their magnetic susceptibility and high adsorption capacity for different types of molecules. A major challenge for separation is to generate selectivity for a target molecule, or for a group of molecules in complex environments such as cell lysates. It is crucial to understand the factors that determine the targets' adsorption behavior in mixtures for triggering intended interactions and selectivity. Here we use a model system containing three molecules, each of them a common representative of the more abundant types of macromolecules in living systems: sodium oleate (SO), a fatty acid; bovine serum albumin (BSA), a protein; and dextran, a polysaccharide. Our results show that (a) the BSA adsorption capacity on the iron oxide material depends markedly on the pH, with the maximum capacity at the pI of the protein (0.39 g g

The demand for purified antibodies is ever-rising. This study presents a nanoparticle-based material for efficient magnetic separation of immunoglobulin G (IgG) with high binding capacity. The char...

Nanoparticles are acquiring an ever increasing role in analytical technologies for enhanced applications such as signalling of hazardous dyes. One challenge for the synthesis of hybrid nanomaterials is to control their shape, size and properties. The colloidal and interfacial properties of initial nanoparticles are decisive for the formation, growth and characteristics of nanohybrids. Our objective is to combine the advantages of iron oxide nanoparticles for magnetic separation with nanoscale gold for a surface enhanced Raman scattering (SERS) effect which could be used e.g. for improved detection of dye molecules. We synthesized iron oxide nanoparticles (∼10 nm) with a high saturation magnetization of around 80 Am2 kg−1 and coupled nanoscale gold to these particles. The focus was set in testing multiple approaches to combine these two materials with the goal of understanding and discussing the effect of the colloidal stability of iron oxide nanoparticles on the properties of the hybrid material. Stability is a seldom addressed issue; however, it plays a critical role for guaranteeing a homogeneous distribution of the gold on the iron oxide surface. We characterized the produced materials with UV/Vis spectroscopy, dynamic light scattering, and transmission electron microscopy, and their capability to enhance Raman signals is investigated. The seed-mediated growth method of oleate and PEG-stabilized magnetic particles yielded the best enhancement of Raman scattering for identification of the dye Rhodamin 6G. This approach can be used to couple gold nanoparticles to other surfaces and microfluidic devices. The presented method might pave the way to further applications in diagnostics or also in environmental approaches and beyond.

Monoclonal antibodies are key molecules in medicine and pharmaceuticals. A potentially crucial drawback for faster advances in research here is their high price due to the extremely expensive antibody purification process, particularly the affinity capture step. Affinity chromatography materials have to demonstrate the high binding capacity and recovery efficiency as well as superior chemical and mechanical stability. Low-cost materials and robust, faster processes would reduce costs and enhance industrial immunoglobulin purification. Therefore, exploring the use of alternative materials is necessary. In this context, we conduct the first comparison of the performance of magnetic nanoparticles with commercially available chromatography resins and magnetic microparticles with regard to immobilizing Protein G ligands and recovering immunoglobulin G (IgG). Simultaneously, we demonstrate the suitability of bare as well as silica-coated and epoxy-functionalized magnetite nanoparticles for this purpose. All materials applied have a similar specific surface area but differ in the nature of their matrix and surface accessibility. The nanoparticles are present as micrometer agglomerates in solution. The highest Protein G density can be observed on the nanoparticles. IgG adsorbs as a multilayer on all materials investigated. However, the recovery of IgG after washing indicates a remaining monolayer, which points to the specificity of the IgG binding to the immobilized Protein G. One important finding is the impact of the ligand-binding stoichiometry (Protein G surface coverage) on IgG recovery, reusability, and the ability to withstand long-term sanitization. Differences in the materials' performances are attributed to mass transfer limitations and steric hindrance. These results demonstrate that nanoparticles represent a promising material for the economical and efficient immobilization of proteins and the affinity purification of antibodies, promoting innovation in downstream processing.

Owing to their high specific surface and low production cost, carbon materials are among the most important adsorption materials. Novel usages, for instance in pharmaceutical applications, challenge existing methods because charged and strongly polar substances need to be adsorbed. Here, we systematically investigate the highly complex adsorption equilibria of organic molecules having multiple protonation states as a function of pH. The adsorption behavior depends on intermolecular interactions within the solution (dissociation equilibria) and between adsorbed molecules on the carbon surface (electrostatic forces). For the model substances maleic acid and phenylalanine, we demonstrate that a custom-made genetic algorithm is able to extract up to nine parameters of a multispecies isotherm from experimental data covering a broad pH-range. The parameters, including adsorption affinities, interaction energies, and maximum loadings were also predicted by molecular dynamics simulations. Both approaches obtained a good qualitative and mostly also quantitative description of the adsorption behavior within a pH-range of 2-12. By combining the determined isotherms with mass balances, the final concentrations and pH-shifts of batch adsorption experiments can be predicted. The developed modeling tools can be easily adapted to other types of pH-dependent, multispecies adsorbates and therefore will help to optimize adsorption-based processes in different fields.

The major interest in nanoparticles as an application platform for biotechnology arises from their high surface-to-volume ratio. Iron oxide nanoparticles (IONPs) are particularly appealing due to their superparamagnetic behavior, which enables bioseparation using external magnetic fields. In order to design advanced biomaterials, improve binding capacities and develop innovative processing solutions, a thorough understanding of the factors governing organic-inorganic binding in solution is critical but has not yet been achieved, given the wide variety of chemical and physical influences. This paper offers a critical review of experimental studies of the interactions between low cost IONPs (bare iron oxides, silica-coated or easily-functionalized surfaces) and the main groups of biomolecules: proteins, lipids, nucleic acids and carbohydrates. Special attention is devoted to the driving forces and interdependencies responsible of interactions at the solid-liquid interface, to the unique structural characteristics of each biomolecular class, and to environmental conditions influencing adsorption. Furthermore, studies focusing on mixtures, which are still rare, but absolutely necessary to understand the biocorona, are also included. This review concludes with a discussion of future work needed to fill the gaps in knowledge of bio-nano interactions, seeking to improve nanoparticles’ targeting capabilities in complex systems, and to open the door for multipurpose recognition and bioseparation processes.

Carbon-based materials are the spearhead of research in multiple fields of nanotechnology. Moreover, their role as stationary phase in chromatography is gaining relevance. We investigate a hybrid material consisting of multiwall carbon nanotubes (CNTs) and superparamagnetic iron oxide nanoparticles (SPIONs) towards its use as a mixed-mode chromatography material. The idea is to immobilize the ion exchange material iron oxide on CNTs as a stable matrix for chromatography processes without a 2 significant pressure drop. Iron oxide nanoparticles are synthesized and used to decorate the CNTs via a co-precipitation route. They bind to the walls of oxidized CNTs, thereby enabling to magnetically separate the hybrid material. This hybrid material is investigated with transmission electron microscopy, magnetometry, X-ray diffraction, X-ray photoelectron and Raman spectroscopy. Moreover, we determine its specific surface area and its wetting behavior. We also demonstrate its applicability as chromatography material for amino acid retention, describing the adsorption and desorption of different amino acids in a complex porous system surrounded by aqueous media. Thus, this material can be used as chromatographic matrix and as a magnetic batch adsorbent material due to the iron oxide nanoparticles. Our work contributes to current research on composite materials. Such materials are necessary for developing novel industrial applications or improving the performance of established processes.

The capture and recovery of valuable products from wastewater and/or bio-broths is one of today’s industrial challenges. Potential-controlled chromatography is a versatile solution to separate charged or polarized target species from aqueous systems. This capacitive flow-through technique combines the advantages of capacitive desalination and ion-exchange chromatography. The separation principle is controlled by system-dependent solid–liquid interaction mechanisms at the potential-controlled interface and characterized by the electrochemical double layer (EDL) formation. We conducted new experiments on the process- and environment-dependent surface effects in aqueous systems to better understand electrically driven processes inside a macroporous multi-walled carbon nanotube electrode. Here, the column and current response are investigated as a function of flow rate, as well as mobile phase pH-value, composition and concentration. Due to the strong water affinity of the material, water ions significantly co-define the EDL. The current response depends particularly on the ion properties and concentration. The use of electrolytes promotes capacitive and faradaic current and diminishes pH-deviation occurring at a negative potential. The effective range of the EDL decreases with increasing shear forces. Finally, a better performance for electrosorbing maleic acid is guaranteed at higher electrolyte concentration and low flow rate. This paper successfully addresses research gaps in potential-dependent effects that occur in multi-walled carbon nanotube electrodes.

Inefficient cell separation is considered to be a critical bottleneck when exploiting microalgal biomass as a renewable resource. Since cell density after cultivation is usually

Inspired by nature, many applications and new materials benefit from the interplay of inorganic materials and biomolecules. A fundamental understanding of complex organic-inorganic interactions would improve the controlled production of nanomaterials and biosensors to the development of biocompatible implants for the human body. Although widely exploited in applications, the interaction of amino acids and peptides with most inorganic surfaces is not fully understood. To date, precisely characterizing complex surfaces of inorganic materials and analyzing surface-biomolecule interactions remain challenging both experimentally and computationally. This article reviews several approaches to characterizing biomolecule-surface interactions and illustrates the advantages and disadvantages of the methods presented. First, we explain how the adsorption mechanism of amino acids/peptides to inorganic surfaces can be determined and how thermodynamic and kinetic process constants can be obtained. Second, we demonstrate how this data can be used to develop models for peptide-surface interactions. The understanding and simulation of such interactions constitute a basis for developing molecules with high affinity binding domains in proteins for bioprocess engineering and future biomedical technologies.

Electrochemically modulated liquid chromatography is a special form of ion exchange chromatography in which the separation process is controlled by applying an electric potential to the stationary phase. This form of chromatography has so far only been applied in research studies. The present study shows that multiwalled carbon nanotubes are an effective resin material for an electrochemically modulated chromatography process. The experiments are carried out in a newly designed column that enables the packing of nanomaterials. We investigate the influence of the applied potential on the retention and elution of maleic acid, determine the dynamic binding capacity, and calculate the utilization degree of the electrical charge in the adsorption process. Moreover, the stability of the resin and the membrane over more than 200 working hours are presented. In addition to the stability, their sturdiness and inexpensive price are important qualities that make multiwalled carbon nanotubes interesting for application as the stationary phase in an electrochemically driven process. The investigated chromatography technique represents a promising separation process for future applications as a preparative step in biotechnology as well as other life science fields.

Iron oxide nanoparticles (IONs) may well represent the most promising magnetic nanostructures for a plethora of applications in health, life and environmental science. IONs are already used in medicine, catalysis and downstream processing of biotechnological products. Since most particles, utilized industrially, need expensive coatings, the application of bare nanoparticles seems economically worthwhile. In this study, three different ION species were synthesized by co-precipitation methods without stabilizing agents and were thoroughly characterized with a multi-analytical approach. We emphasize the importance of the particle characterization as transitions of the ION polymorphs into each other are possible as well as merging of distinct properties. The particle sizes, which here range from 10 to 30 nm, and the magnetic properties of IONs are crucial for the further application. The adsorption behavior of the enzyme cellulase (CEL) as a model protein is investigated on the different IONs in order to gain deeper insights into bio-nano interactions to different surface sites, charges, curvatures and morphologies, as given by the three applied adsorber materials. The protein-particle interactions are driven by electrostatic and hydrophobic forces in the case of CEL. The CEL adsorption follows a Langmuir behavior and does not exceed maximum loads of around 0.6 g g−1. IR spectroscopy gives insights into the orientation of bound CEL and indicates a stronger affinity for the β-sheet tertiary structure content while a higher load can be reached with a higher α-helix content.

Magnetic separation is a promising alternative to conventional methods in downstream processing. This can facilitate easier handling, fewer processing steps, and more sustainable processes. Target materials can be extracted directly from crude cell lysates in a single step by magnetic nanoadsorbents with high-gradient magnetic fishing (HGMF). Additionally, the use of hazardous consumables for reducing downstream processing steps can be avoided. Here, we present proof of principle of one-step magnetic fishing from crude Escherichia coli cell lysate of a green fluorescent protein (GFP) with an attached hexahistidine (His6)-tag, which is used as the model target molecule. The focus of this investigation is the upscale to a liter scale magnetic fishing process in which a purity of 91% GFP can be achieved in a single purification step from cleared cell lysate. The binding through the His6-tag can be demonstrated, since no significant binding of nontagged GFP toward bare iron oxide nanoparticles (BIONs) can be observed. Nonfunctionalized BIONs with primary particle diameters of around 12 nm, as used in the process, can be produced with a simple and low-cost coprecipitation synthesis. Thus, HGMF with BIONs might pave the way for a new and greener era of downstream processing.

Potential-controlled tensiometry is a voltage-induced method which enables measuring the contact angle between a powder bed and a liquid medium through the capillary rise method. This analytical tool provides a fine-grained technique for understanding wetting behavior of powders as well as solid surfaces in connection with the application of an electrical potential. In this work, the powder bed was brought into contact with an aluminum rod connected to a portable lightweight DAC-module (digital to analog converter) powered by a lithium-polymer battery (LiPo). The presented analytical device can be charged up to ±1000 mV. Both the power source and the DAC-module are lightweight in order to be conveniently attached to a force tensiometer without incorporating complex wiring. In this setup, we tested multiwall carbon nanotubes (MWCNT) and glassy carbon particles. An influence of the potential on the wetting behavior of glassy carbon particles is observed which demonstrates the working principle of the device. Surprisingly, no significant effect of the potential on the wetting behavior of MWCNT is indicated in the range studied. This technique can be a valuable tool to analyze the effect of changing surface properties applying electrical gradients on materials.

Owing to their chemical and magnetic properties, magnetite nanoparticles are an interesting adsorbing material for biomolecules. The understanding of the interactions of simple biomolecules with inorganic nanoparticles is an important approach for research on the bio-nano interface and can constitute the fundamentals to manifold applications in biotechnology, medicine and catalysis. The aim of the work presented here is to compare the interaction of seven different amino acids (l-alanine, l-cysteine, l-glutamic acid, glycine, l-histidine, l-lysine, and l-serine) with magnetite nanoparticles in a colloidal system at pH 6. We investigate the influence of the side chain on the adsorption at a magnetite–water interface with incubation experiments. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and simultaneous thermal analysis (STA) reveal deeper insights into the interactions of amino acids with magnetite nanoparticles. The amino acids...

Superparamagnetic nanoparticles have recently gained much attention due to their broad range of applicability including medical in vivo technologies, sensors, and as supports for catalysts. As magnetic affinity materials, they can be utilized for the development of new purification strategies for pharmaceuticals and other target molecules from crude lysates. Here, a short peptide tag based on a glutamate sequence is introduced and the adsorption of pure protein as well as protein from crude cell lysate at different conditions is demonstrated. Fused to a model protein this tag can be used to recognize and purify this protein from a fermentation broth by bare iron oxide nanoparticles (BIONs). Binding of up to 0.2 g protein per g nanoparticles can be achieved and recovered easily by switching to a citrate buffered system. For a deeper understanding of the separation process, the aggregation and agglomeration of the nanoparticle protein systems were monitored for binding and elution steps. Furthermore, an upscaling of the process to the liter scale and the separation of a green fluorescent protein (GFP) containing the affinity tag to purities of 70% from Escherichia coli fermentation broth was possible in a one step process by means of high gradient magnetic separation (HGMS).

Magnetic metal oxide nanoparticles demonstrate great applicability in several fields such as biotechnology, medicine and catalysis. A stable, magnetic and low-cost material, nanoscale magnetite, is an interesting adsorbent for protein purification. Downstream processing can account for up to 80% of the total production costs in biotechnological production. As such, the development of new innovative separation methods can be regarded as highly profitable. While short peptide sequences can be used as specific affinity tags for functionalised adsorber surfaces, they need expensive affinity ligands on the particle surface for adsorption. In order to identify peptide tags for several non-functionalised inorganic surfaces, different binding conditions to iron oxide nanoparticles are evaluated. Therefore, magnetite nanoparticles in a range of 5–20 nm were synthesised with a co-precipitation method. Zeta potential measurements indicated an amphiphilic surface with an isoelectric point in the neutral pH region. Glutamic acid-based homo-peptides were used as affinity peptides for the magnetite nanoparticles. We demonstrate a dependence of the binding affinity of the peptides on pH and buffer ions in two different experimental set-ups. The nature of surface coordination for glutamic acid-based peptides can be demonstrated with different spectroscopic approaches such as infrared spectroscopy (IR), Raman spectroscopy and circular dichroism spectroscopy (CD). We want to emphasise the importance of physicochemical properties such as surface energy, polarity, morphology and charge. These parameters, which are dependent on the environmental conditions, play a crucial role in peptide interactions with iron oxide surfaces. The understanding of the adsorption of simple biomolecules on nanoscale metal oxide surfaces also represents the key to the even more complex interactions of proteins at the bio-nano interface. From the identification of interaction patterns and an understanding of the adsorption of these peptides, the up-scaling to tagged model proteins facilitates the possibility of an industrial magnetic separation process and might therefore reduce time and costs in purification processes.

Iron oxide nanoparticles represent a promising low-cost environmentally-friendly material for multiple applications. Especially hematite (α-Fe2O3) nanoparticles demonstrate great possibilities in energy storage and photoelectrochemistry. A hydrothermal one-pot synthesis can be used to synthesise hematite nanoparticles. Here, the particle formation, nucleation and growth of iron oxide nanoparticles using a FeCl3 precursor over time is monitored. The formation of 6-line ferrihydrite seeds of 2–8 nm which grow with reaction time and form clusters followed by a phase transition to ~15 nm hematite particles can be observed with ex situ X-ray diffraction (XRD), transmission electron microscopy (TEM), Raman and UV/Vis spectroscopy. These particles grow with reaction time leading to 40 nm particles after 6 hours. The changes in plasmon and electron transition patterns, observed upon particle transition and growth lead to the possibility of tuning the photoelectrochemical properties. Catalytic activity of the hematite nanoparticles can be proven with visible light irradiation and the use of silver nitrate as scavenger material. The generation of elementary silver is dependent on the particle size of iron oxide nanoparticles while only slight changes can be observed in the oxygen generation. Low-cost nanoscale hematite, offers a range of future applications for artificial photosynthesis.

Iron oxide nanoparticles are of great scientific interest due to their huge versatility of applications. The oxidation process of magnetite to maghemite is difficult to monitor as both iron oxide polymorphs possess connatural chemical properties. Especially the surface composition and reactivity of these nanosystems, which are most relevant for interactions with their environment, are not completely understood. Here, the oxidation of magnetite is investigated under mild and harsh conditions in order to understand the oxidation behaviour and the chemical stability of transition forms. Therefore, the oxidation process, is investigated with Raman, Mossbauer and X-ray photoelectron spectroscopy as well as X-ray diffraction and magnetometry. The multi-analytical approach allows new insights into surface composition and rearrangement according to respective different depth profiles. For both conditions investigated, the ferrous iron components are oxidised prior to structural changes in the Fe–O vibrations and crystal structure. The process starts from the outer layers and is acid catalysed. Oxidation leads to a decrease of magnetisation which still remains higher than 54 emu g−1. The charge and surface reactivity can be affected by the different oxidation methods and the irreversible adsorption of acid molecules. Biocompatibility and catalytic properties of iron oxide nanoparticles open doors to future applications.

Magnetic nanoparticles (MNP) are intensively investigated for applications in nanomedicine, catalysis and biotechnology, where their interaction with peptides and proteins plays an important role. However, the characterisation of the interaction of individual amino acids with MNP remains challenging. Here, we classify the affinity of 20 amino acid homo-hexamers to unmodified iron oxide nanoparticles using peptide arrays in a variety of conditions as a basis to identify and rationally design selectively binding peptides. The choice of buffer system is shown to strongly influence the availability of peptide binding sites on the MNP surface. We find that under certain buffer conditions peptides of different charges can bind the MNP and that the relative strength of the interactions can be modulated by changing the buffer. We further present a model for the competition between the buffer and the MNP’s electrostatically binding to the adsorption sites. Thereby, we demonstrate that the charge distribution on the surface can be used to correlate the binding of positively and negatively charged peptides to the MNP. This analysis enables us to engineer the binding of MNP on peptides and contribute to better understand the bio-nano interactions, a step towards the design of affinity tags for advanced biomaterials.

Electrochemically modulated liquid chromatography is a special form of ion exchange chromatography in which the separation process is controlled by applying an electric potential to the stationary phase. This form of chromatography has so far only been applied in research studies. The present study shows that multiwalled carbon nanotubes are an effective resin material for an electrochemically modulated chromatography process. The experiments are carried out in a newly designed column that enables the packing of nanomaterials. We investigate the influence of the applied potential on the retention and elution of maleic acid, determine the dynamic binding capacity, and calculate the utilization degree of the electrical charge in the adsorption process. Moreover, the stability of the resin and the membrane over more than 200 working hours are presented. In addition to the stability, their sturdiness and inexpensive price are important qualities that make multiwalled carbon nanotubes interesting for application as the stationary phase in an electrochemically driven process. The investigated chromatography technique represents a promising separation process for future applications as a preparative step in biotechnology as well as other life science fields.

Coating magnetic nanoparticles (MNPs) with sodium oleate (SO) is known to be an excellent method to create biocompatible, stable colloids with a narrow size distribution. However, the mechanism of oleate adsorption on the MNP surface in aqueous systems, as well as its influence on colloidal stability, is not yet fully understood. In this context, we present here a physico-chemical study to provide a deeper understanding of surfactant interaction mechanisms with nanoparticles. We examined the effect of temperature and the SO/MNP ratio (w/w) on the adsorption process in water and observed the existence of a maximum for the adsorbed oleate amount at lower temperatures, whereas at higher temperatures, the isotherm can be adapted to the Langmuir model with constant capacity after saturation. The oleate load on the MNP surface was quantified using reversed-phase high-performance liquid chromatography measurements of samples in solution. The thermogravimetric analyses of the solid residues together with infrared spectroscopy analyses indicate a bilayer-similar structure at the MNP/water interface even for low oleate loads. The oleate interacts with the iron oxide surface through a bidentate coordination of the carboxyl group. Zeta potential measurements demonstrate the high stability of the coated system. The maximal oleate load per unit mass of MNPs reaches approximately 0.35 goleate g MNP −1 .