Search

Abstract Liquid‐solid and liquid‐liquid phase separation (PS) drives the formation of functional and disease‐associated biological assemblies. Principles of phase equilibrium are here employed to derive a general kinetic solution that predicts the evolution of the mass and size of biological assemblies. Thermodynamically, protein PS is determined by two measurable concentration limits: the saturation concentration and the critical solubility. Due to surface tension effects, the critical solubility can be higher than the saturation concentration for small, curved nuclei. Kinetically, PS is characterized by the primary nucleation rate constant and a combined rate constant accounting for growth and secondary nucleation. It is demonstrated that the formation of a limited number of large condensates is possible without active mechanisms of size control and in the absence of coalescence phenomena. The exact analytical solution can be used to interrogate how the elementary steps of PS are affected by candidate drugs.

Van Lindt, Bratek-Skicki et al. show that at carefully selected pH values, proteins can be kept in solution and their LLPS can then be induced by a jump to native pH. This presents a generic method to study the full kinetic trajectory of LLPS under near native conditions that can be easily controlled, providing a platform for the characterization of physiologically relevant phase-separation behaviour of diverse proteins.

The rapidly-growing field of polymer stimuli-responsive materials (smart materials) resulted in many advances in designing and developing multi-functional systems, novel architectures, and new synthetic routes for many different applications. The smart polymer materials with a wide range of stimuli, such as pH, light, temperature, enzyme, redox, and electric and magnetic fields play an important role in many applications in many disciplines, including nanotechnology, biochemistry, medicine, materials science, polymer science, engineering, etc. This review aims to provide an introduction to stimuli-responsive polymeric materials, their preparation methods, and to discuss the design of various stimulus-responsive materials that are able to provide smart self-healing surfaces, sensors, actuators, and drug delivery systems in response to particular stimuli.

Aggregates of TAR DNA-binding protein (TDP-43) are a hallmark of several neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). Although TDP-43 aggregates are an undisputed pathological species at the end stage of these diseases, the molecular changes underlying the initiation of aggregation are not fully understood. The aim of this study was to investigate how phase separation affects self-aggregation and aggregation seeded by pre-formed aggregates of either the low-complexity domain (LCD) or its short aggregation-promoting regions (APRs). By systematically varying the physicochemical conditions, we observed that liquid–liquid phase separation (LLPS) promotes spontaneous aggregation. However, we noticed less efficient seeded aggregation in phase separating conditions. By analyzing a broad range of conditions using the Hofmeister series of buffers, we confirmed that stabilizing hydrophobic interactions prevail over destabilizing electrostatic forces. RNA affected the cooperativity between LLPS and aggregation in a “reentrant” fashion, having the strongest positive effect at intermediate concentrations. Altogether, we conclude that conditions which favor LLPS enhance the subsequent aggregation of the TDP-43 LCD with complex dependence, but also negatively affect seeding kinetics.

A Correction to this paper has been published: https://doi.org/10.1038/s42003-021-01853-4.

Controlled deposition of nanoparticles and bioparticles is necessary for their separation and purification by chromatography, filtration, food emulsion and foam stabilization, etc. Compared to numerous experimental techniques used to quantify bioparticle deposition kinetics, the quartz crystal microbalance (QCM) method is advantageous because it enables real time measurements under different transport conditions with high precision. Because of its versatility and the deceptive simplicity of measurements, this technique is used in a plethora of investigations involving nanoparticles, macroions, proteins, viruses, bacteria and cells. However, in contrast to the robustness of the measurements, theoretical interpretations of QCM measurements for a particle-like load is complicated because the primary signals (the oscillation frequency and the band width shifts) depend on the force exerted on the sensor rather than on the particle mass. Therefore, it is postulated that a proper interpretation of the QCM data requires a reliable theoretical framework furnishing reference results for well-defined systems. Providing such results is a primary motivation of this work where the kinetics of particle deposition under diffusion and flow conditions is discussed. Expressions for calculating the deposition rates and the maximum coverage are presented. Theoretical results describing the QCM response to a heterogeneous load are discussed, which enables a quantitative interpretation of experimental data obtained for nanoparticles and bioparticles comprising viruses and protein molecules.

Liquid‐solid and liquid‐liquid phase separation (PS) drives the formation of functional and disease‐associated biological assemblies. Principles of phase equilibrium are here employed to derive a general kinetic solution that predicts the evolution of the mass and size of biological assemblies. Thermodynamically, protein PS is determined by two measurable concentration limits: the saturation concentration and the critical solubility. Due to surface tension effects, the critical solubility can be higher than the saturation concentration for small, curved nuclei. Kinetically, PS is characterized by the primary nucleation rate constant and a combined rate constant accounting for growth and secondary nucleation. It is demonstrated that the formation of a limited number of large condensates is possible without active mechanisms of size control and in the absence of coalescence phenomena. The exact analytical solution can be used to interrogate how the elementary steps of PS are affected by candidate drugs. [ABSTRACT FROM AUTHOR]

Manipulation of high-density materials, such as crystals and liquid condensates, is of great importance for many applications, including serial crystallography, structural and molecular biology, chemistry, and medicine. In this work, we describe an acoustic technique to focus and harvest flowing crystals and liquid condensates. Moreover, we show, based on numerical simulations, that the acoustic waves can be used for size-based particle (crystals, droplets, etc.) separation. This is an essential technological step in biological research, medical applications, and industrial processes. The presented technology offers high precision, biocompatibility, ease of use and additionally, is non-invasive and inexpensive. With the recent advent of X-ray Free Electron Laser (XFEL) technology and the associated enormous importance of a thin jet of crystals, this technology might pave the way to a novel type of XFEL injector.

Adsorption of fibrinogen on positively charged microspheres was theoretically and experimentally studied. The structure of monolayers and the maximum coverage were determined by applying the experimental measurements at pH = 3.5 and 9.7 for NaCl concentration in the range of 10^{-3} - 0.15 M. The maximum coverage of fibrinogen on latex particles was precisely determined by the AFM method. Unexpectedly, at pH = 3.5, where both fibrinogen molecule and the latex particles were positively charged, the maximum coverage varied between 0.9 mg m^{-2} and 1.1 mg m^{-2} for 10^{-2} and 0.15 M NaCl, respectively. On the other hand, at pH = 9.7, the maximum coverage of fibrinogen was larger, varying between 1.8 mg m^{-2} and 3.4 mg m^{-2} for 10^{-2} and 0.15 M NaCl, respectively. The experimental results were quantitatively interpreted by the numerical simulations.

Motivation Proteins able to undergo liquid–liquid phase separation (LLPS) in vivo and in vitro are drawing a lot of interest, due to their functional relevance for cell life. Nevertheless, the proteome-scale experimental screening of these proteins seems unfeasible, because besides being expensive and time-consuming, LLPS is heavily influenced by multiple environmental conditions such as concentration, pH and temperature, thus requiring a combinatorial number of experiments for each protein. Results To overcome this problem, we propose a neural network model able to predict the LLPS behavior of proteins given specified experimental conditions, effectively predicting the outcome of in vitro experiments. Our model can be used to rapidly screen proteins and experimental conditions searching for LLPS, thus reducing the search space that needs to be covered experimentally. We experimentally validate Droppler's prediction on the TAR DNA-binding protein in different experimental conditions, showing the consistency of its predictions. Availability and implementation A python implementation of Droppler is available at https://bitbucket.org/grogdrinker/droppler Supplementary information Supplementary data are available at Bioinformatics online. [ABSTRACT FROM AUTHOR]

Cellular organelles that lack a surrounding lipid bilayer, such as the nucleolus and stress granule, represent a newly recognized, general paradigm of cellular organization. The formation of such biomolecular condensates that include 'membraneless organelles' (MLOs) by liquid–liquid phase separation (LLPS) has been in the focus of a surge of recent studies. Through a combination of in vitro and in vivo approaches, thousands of potential phase‐separating proteins have been identified, and it was found that different cellular MLOs share many common components. These perplexing observations raise the question of how cells regulate the timing and specificity of LLPS, and ensure that different MLOs form and disperse at the right moment and cellular location and can preserve their identity and physical separation. This guide gives an overview of basic regulatory mechanisms, which manifest through the action of intrinsic regulatory elements, alternative splicing, post‐translational modifications, and a broad range of phase‐separating partners. We also elaborate on the cellular integration of these different mechanisms and highlight how complex regulation can orchestrate the parallel functioning of a dozen or so different MLOs in the cell. [ABSTRACT FROM AUTHOR]

Proteins at interfaces are a key for many applications in the biomedical field, in biotechnologies, in biocatalysis, in food industry, etc. The development of surface layers that allow to control and manipulate proteins is thus highly desired. In previous works, we have shown that mixed polymer brushes combining the protein-repellent properties of poly(ethylene oxide) (PEO) and the stimuli-responsive adsorption behavior of poly(acrylic acid) (PAA) could be synthesized and used to achieve switchable protein adsorption. With the present work, we bring more insight into the rational design of such smart thin films by unravelling the role of PEO on the adsorption/desorption of proteins. The PEO content of the mixed PEO/PAA brushes was regulated, on the one hand, by using PEO with different molar masses and, on the other hand, by varying the ratio of PEO and PAA in the solutions used to synthesize the brushes. The influence of ionic strength on the protein adsorption behavior was also further examined. The behavior of three proteins--human serum albumin, lysozyme, and human fibrinogen, which have very different size, shape, and isoelectric point--was investigated. X-ray photoelectron spectroscopy, quartz crystal microbalance, atomic force microscopy, and streaming potential measurements were used to characterize the mixed polymer brushes and, in particular, to estimate the fraction of each polymer within the brushes. Protein adsorption and desorption conditions were selected based on previous studies. While brushes with a lower PEO content allowed the higher protein adsorption to occur, fully reversible adsorption could only be achieved when the PEO surface density was at least 25 PEO units per nm². Taken together, the results increase the ability to finely tune protein adsorption, especially with temporal control. This opens up possibilities of applications in biosensor design, separation technologies, nanotransport, etc. [ABSTRACT FROM AUTHOR]

Adsorption of fibrinogen on positively charged microspheres was theoretically and experimentally studied. The structure of monolayers and the maximum coverage were determined by applying the experimental measurements at pH = 3.5 and 9.7 for NaCl concentration in the range of 10-3 -0.15 M. The maximum coverage of fibrinogen on latex particles was precisely determined by the AFM method. Unexpectedly, at pH = 3.5, where both fibrinogen molecule and the latex particles were positively charged, the maximum coverage varied between 0.9 mg m-2 and 1.1 mg m-2 for 10-2 and 0.15 M NaCl, respectively. On the other hand, at pH = 9.7, the maximum coverage of fibrinogen was larger, varying between 1.8 mgm-2 and 3.4 mgm-2 for 10-2 and 0.15 M NaCl, respectively. The experimental results were quantitatively interpreted by the numerical simulations. [ABSTRACT FROM AUTHOR]

Fibrinogen (Fb) adsorption on positivelycharged latex particles(average diameter of 800 nm) was studied using the microelectrophoreticand the concentration depletion methods based on AFM imaging. Monolayerson latex were adsorbed from diluted bulk solutions at pH 7.4 and anionic strength in the range of 10–3to 0.15 M wherefibrinogen molecules exhibited an average negative charge. The electrophoreticmobility of the latex after controlled fibrinogen adsorption was systematicallymeasured. A monotonic decrease in the electrophoretic mobility offibrinogen-covered latex was observed for all ionic strengths. Theresults of these experiments were interpreted according to the three-dimensionalelectrokinetic model. It was also determined using the concentrationdepletion method that fibrinogen adsorption was irreversible and themaximum coverage was equal to 0.6 mg m–2for ionicstrength 10–3M and 1.3 mg m–2for ionic strength 0.15 M. The increase of the maximum coveragewas confirmed by theoretical modeling based on the random sequentialadsorption approach. Paradoxically, the maximum coverage of fibrinogenon positively charged latex particles was more than two times lowerthan the maximum coverage obtained for negative latex particles (3.2mg m–2) at pH 7.4 and ionic strength of 0.15 M.This was interpreted as a result of the side-on adsorption of fibrinogenmolecules with their negatively charged core attached to the positivelycharged latex surface. The stability and acid base properties of fibrinogenmonolayers on latex were also determined in pH cycling experimentswhere it was observed that there were no irreversible conformationalchanges in the fibrinogen monolayers. Additionally, the zeta potentialof monolayers was more positive than the zeta potential of fibrinogenin the bulk, which proves a heterogeneous charge distribution. Theseexperimental data reveal a new, side-on adsorption mechanism of fibrinogenon positively charged surfaces and confirmed the decisive role ofelectrostatic interactions in this process. [ABSTRACT FROM AUTHOR]

The adsorption of fibrinogen on polystyrene latex particles was studied using the concentration depletion method combined with the AFM detection of residual protein after adsorption. Measurements were carried out for a pH range of 3.5–11 and an ionic strength range of 10–3–0.15 M NaCl. First, the bulk physicochemical properties of fibrinogen and the latex particle suspension were characterized for this range of pH and ionic strength. The zeta potential and the number of uncompensated (electrokinetic) charges on the protein were determined from microelectrophoretic measurements. It was revealed that fibrinogen molecules exhibited amphoteric characteristics, being on average positively charged for pH 5.8 and a higher ionic strength of 0.15 M. It was also shown that in the latter case, variations in the zeta potential with the protein coverage could be adequately described in terms of the electrokinetic model, previously formulated for planar substrate adsorption. On the basis of these experimental data, an efficient procedure of preparing fibrinogen-covered latex particles of controlled monolayer structure and coverage was envisaged. [ABSTRACT FROM AUTHOR]

Aggregates of TAR DNA-binding protein (TDP-43) are a hallmark of several neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). Although TDP-43 aggregates are an undisputed pathological species at the end stage of these diseases, the molecular changes underlying the initiation of aggregation are not fully understood. The aim of this study was to investigate how phase separation affects self-aggregation and aggregation seeded by pre-formed aggregates of either the low-complexity domain (LCD) or its short aggregation-promoting regions (APRs). By systematically varying the physicochemical conditions, we observed that liquid–liquid phase separation (LLPS) promotes spontaneous aggregation. However, we noticed less efficient seeded aggregation in phase separating conditions. By analyzing a broad range of conditions using the Hofmeister series of buffers, we confirmed that stabilizing hydrophobic interactions prevail over destabilizing electrostatic forces. RNA affected the cooperativity between LLPS and aggregation in a "reentrant" fashion, having the strongest positive effect at intermediate concentrations. Altogether, we conclude that conditions which favor LLPS enhance the subsequent aggregation of the TDP-43 LCD with complex dependence, but also negatively affect seeding kinetics. [ABSTRACT FROM AUTHOR]

Controlled deposition of nanoparticles and bioparticles is necessary for their separation and purification by chromatography, filtration, food emulsion and foam stabilization, etc. Compared to numerous experimental techniques used to quantify bioparticle deposition kinetics, the quartz crystal microbalance (QCM) method is advantageous because it enables real time measurements under different transport conditions with high precision. Because of its versatility and the deceptive simplicity of measurements, this technique is used in a plethora of investigations involving nanoparticles, macroions, proteins, viruses, bacteria and cells. However, in contrast to the robustness of the measurements, theoretical interpretations of QCM measurements for a particle-like load is complicated because the primary signals (the oscillation frequency and the band width shifts) depend on the force exerted on the sensor rather than on the particle mass. Therefore, it is postulated that a proper interpretation of the QCM data requires a reliable theoretical framework furnishing reference results for well-defined systems. Providing such results is a primary motivation of this work where the kinetics of particle deposition under diffusion and flow conditions is discussed. Expressions for calculating the deposition rates and the maximum coverage are presented. Theoretical results describing the QCM response to a heterogeneous load are discussed, which enables a quantitative interpretation of experimental data obtained for nanoparticles and bioparticles comprising viruses and protein molecules. [ABSTRACT FROM AUTHOR]

Protein adsorption on solid surfaces provides either beneficial or adverse outcomes, depending on the application. Therefore, the desire to predict, control, and regulate protein adsorption on different surfaces is a major concern in the field of biomaterials. The most widely used surface modification approach to prevent or limit protein adsorption is based on the use of poly (ethylene oxide) (PEO). On the other hand, the amount of protein adsorbed on poly(2-(dimethylamine)ethyl methacrylate) (PDMAEMA) coatings can be regulated by the pH and ionic strength of the medium. In this work, ultra-thin PEO/PDMAEMA coatings were designed from solutions with different ratios of PEO to PDMAEMA, and different molar masses of PEO, to reversibly adsorb and desorb human serum albumin (HSA), human fibrinogen (Fb), lysozyme (Lys), and avidine (Av), four very different proteins in terms of size, shape, and isoelectric points. X-ray photoelectron spectroscopy (XPS), quartz crystal microbalance (QCM), and atomic force microscopy (AFM) were used to characterize the mixed polymer coatings, revealing the presence of both polymers in the layers, in variable proportions according to the chosen parameters. Protein adsorption at pH 7.4 and salt concentrations of 10−3 M was monitored by QCM. Lys and Av did not adsorb on the homo-coatings and the mixed coatings. The amount of HSA and Fb adsorbed decreased with increasing the PEO ratio or its molar mass in a grafting solution. It was demonstrated that HSA and Fb, which were adsorbed at pH 7.4 and at an ionic strength of 10−3 M, can be fully desorbed by rinsing with a sodium chloride solution at pH 9.0 and ionic strength 0.15 M from the mixed PEO5/PDMAEMA coatings with PEO/PDMAEMA mass ratios of 70/30, and 50/50, respectively. The results demonstrate that mixed PEO/PDMAEMA coatings allow protein adsorption to be finely tuned on solid surfaces. [ABSTRACT FROM AUTHOR]

Manipulation of high-density materials, such as crystals and liquid condensates, is of great importance for many applications, including serial crystallography, structural and molecular biology, chemistry, and medicine. In this work, we describe an acoustic technique to focus and harvest flowing crystals and liquid condensates. Moreover, we show, based on numerical simulations, that the acoustic waves can be used for size-based particle (crystals, droplets, etc.) separation. This is an essential technological step in biological research, medical applications, and industrial processes. The presented technology offers high precision, biocompatibility, ease of use and additionally, is non-invasive and inexpensive. With the recent advent of X-ray Free Electron Laser (XFEL) technology and the associated enormous importance of a thin jet of crystals, this technology might pave the way to a novel type of XFEL injector. [ABSTRACT FROM AUTHOR]

Review: 100 refs. [ABSTRACT FROM AUTHOR]

Layer-by-layer (LbL) assembly is an attractive method for protein immobilization at interfaces, a much wanted step for biotechnologies and biomedicine. Integrating proteins in LbL thin films is however very challenging due to their low conformational entropy, heterogeneous spatial distribution of charges, and polyampholyte nature. Protein-polyelectrolyte complexes (PPCs) are promising building blocks for LbL construction owing to their standardized charge and polyelectrolyte (PE) corona. In this work, lysozyme was complexed with poly(styrenesulfonate) (PSS) at different ionic strengths and pH values. The PPCs size and electrical properties were investigated, and the forces driving complexation were elucidated, in the light of computations of polyelectrolyte conformation, with a view to further unravel LbL construction mechanisms. Quartz crystal microbalance and atomic force microscopy were used to monitor the integration of PPCs compared to the one of bare protein molecules in LbL assemblies, and colorimetric assays were performed to determine the protein amount in the thin films. Layers built with PPCs show higher protein contents and hydration levels. Very importantly, the results also show that LbL construction with PPCs mainly relies on standard PE-PE interactions, independent of the charge state of the protein, in contrast to classical bare protein assembly with PEs. This considerably simplifies the incorporation of proteins in multilayers, which will be beneficial for biosensing, heterogeneous biocatalysis, biotechnologies, and medical applications that require active proteins at interfaces.

Human fibrinogen adsorption on negatively charged latex particles was investigated using the electrophoretic and the concentration depletion methods. Measurements were conducted at pH 7.4 and in the range of ionic strength of 10(-3) - 0.15 M NaCl. Firstly, the bulk physicochemical properties of fibrinogen were characterized. The zeta potential and the uncompensated (electrokinetic) charges of the protein were determined from the electrophoretic measurements. Next, systematic experiments were performed to determine the dependencies of the electrophoretic mobility of latex on the amount of adsorbed protein. Electrophoretic mobility increased significantly upon fibrinogen adsorption that was proven irreversible. The maximum coverage of fibrinogen on latex particles determined via the concentration depletion method varied between 1.9 mg m(-2) and 3.2 mg m(-2) for 10(-3) and 0.15 M NaCl, respectively. The changes in the maximum coverage were interpreted as due to electrostatic repulsion among adsorbed protein molecules. Additionally, the stability of latex covered by fibrinogen was determined. It was proven that cyclic changes of ionic strength from 10(-3) to 0.15 M NaCl did not change the electrophoretic mobility. Based on these observations, it was concluded that there were no conformational changes within adsorbed fibrinogen molecules. The experimental data, allowed one to elaborate a robust procedure of preparing latex particles covered by fibrinogen of designed coverage and molecule distribution.

The aim of this work was to critically review recent results pertinent to fibrinogen adsorption at solid/electrolyte interfaces with the emphasis focused on a quantitative analysis of these processes in terms of the electrostatic interactions. Accordingly, in the first part, the primary chemical structure of fibrinogen is analyzed. Physicochemical data pertinent to the bulk properties derived from hydrodynamic, dynamic light scattering and micro-electrophoretic measurements aided by theoretical modeling are discussed. Possible conformations and the effective charge distribution over the fibrinogen molecule for various pH an ionic strength are defined, especially the semi-collapsed conformation prevailing at physiological conditions. Adsorption kinetics of fibrinogen at hydrophilic and hydrophobic (polymer modified) substrates determined by various techniques is described. Adsorption at polymeric carrier particles, pertinent to immunological assays, studied in terms of electrokinetic and concentration depletion methods, are also considered. The reversibility of adsorption, fibrinogen molecule orientations and maximum coverages are thoroughly discussed. The stability of fibrinogen monolayers formed at these carrier particles in respect to pH and ionic strength cyclic changes is also discussed. In the final section interactions and deposition of model colloid particles on fibrinogen monolayers are analyzed which allows one to derive valuable information about molecule orientations. Based on the physicochemical data, adsorption kinetics and colloid particle deposition measurements, probable adsorption mechanisms of fibrinogen on solid/electrolyte interfaces are defined.

The adsorption of human serum fibrinogen on polystyrene latex particles was studied using the microelectrophoretic and concentration depletion methods. Measurements were carried out for pH 3.5 and an ionic strength range of 10(-3) to 0.15 M NaCl. The electrophoretic mobility of latex was determined as a function of the amount of adsorbed fibrinogen (surface concentration). A monotonic increase in the electrophoretic mobility (zeta potential) of the latex was observed, indicating a significant adsorption of fibrinogen on latex for all ionic strengths. No changes in the latex mobility were observed for prolonged time periods, suggesting the irreversibility of fibrinogen adsorption. The maximum coverage of fibrinogen on latex particles was precisely determined using the depletion method. The residual protein concentration after making contact with latex particles was determined by electrokinetic measurements and AFM imaging where the surface coverage of fibrinogen on mica was quantitatively determined. The maximum fibrinogen coverage increased monotonically with ionic strength from 1.8 mg m(-2) for 10(-3) M NaCl to 3.6 mg m(-2) for 0.15 M NaCl. The increase in the maximum coverage was interpreted in terms of the reduced electrostatic repulsion among adsorbed fibrinogen molecules. The experimental data agree with theoretical simulations made by assuming a 3D unoriented adsorption of fibrinogen. The stability of fibrinogen monolayers on latex was also determined in ionic strength cycling experiments. It was revealed that cyclic variations in NaCl concentration between 10(-3) and 0.15 M induced no changes in the latex electrophoretic mobility, suggesting that there were no irreversible molecule orientation changes in the monolayers. On the basis of these experimental data, a robust procedure of preparing fibrinogen monolayers on latex particles of well-controlled coverage was proposed.

The adsorption of fibrinogen on polystyrene latex particles was studied using the concentration depletion method combined with the AFM detection of residual protein after adsorption. Measurements were carried out for a pH range of 3.5-11 and an ionic strength range of 10(-3)-0.15 M NaCl. First, the bulk physicochemical properties of fibrinogen and the latex particle suspension were characterized for this range of pH and ionic strength. The zeta potential and the number of uncompensated (electrokinetic) charges on the protein were determined from microelectrophoretic measurements. It was revealed that fibrinogen molecules exhibited amphoteric characteristics, being on average positively charged for pH <5.8 (isolectric point) and negative otherwise. However, the latex particles did not show any isoelectric point, remaining strongly negative for this pH range. Afterward, systematic measurements of the electrophoretic mobility of fibrinogen-covered latex were carried out as a function of the amount of adsorbed protein, expressed as the surface concentration. A monotonic increase in the electrophoretic mobility (zeta potential) of the latex was observed in all cases, indicating a significant adsorption of fibrinogen on latex for pH below 11. It was also proven that fibrinogen adsorption was irreversible, with the maximum surface concentration varying between 2.5 and 5 × 10(3) μm(-2) (weight concentration of a bare molecule was 1.4 to 2.8 mg m(-2)). These measurements revealed two main adsorption mechanisms of fibrinogen: (i) the unoriented (random) mechanism prevailing for lower ionic strength, where adsorbing molecules significantly penetrate the fuzzy polymeric layer on the latex core and (ii) the side-on adsorption mechanism prevailing for pH > 5.8 and a higher ionic strength of 0.15 M. It was also shown that in the latter case, variations in the zeta potential with the protein coverage could be adequately described in terms of the electrokinetic model, previously formulated for planar substrate adsorption. On the basis of these experimental data, an efficient procedure of preparing fibrinogen-covered latex particles of controlled monolayer structure and coverage was envisaged.