Protein-polymer Interactions Research Articles

Tuning Polymer-Protein Interaction with Salt

Biological nanopores are known to interact with synthetic and biological polymers, enabling their use in label-free single-molecule analytical tasks such as sequencing and/or mass discrimination. The latter, called nanopore-based single molecule mass spectrometry (Np-SMMS) has, to date, only been shown for one synthetic polymer, poly(ethyleneglycol) (PEG). It is based on the fact that binding of PEG inside the pore gives rise to blocks of ionic current, with the degree of block being detectably different for PEG molecules differing in size by only one monomer1,2.In order to extend the range of application of Np-SMMS, and to obtain information on the mechanism underying mass sensitivity of polymer-induced pore block, we have begun to test the interaction of other neutral polymers with biological nanoores. Here, we report on poly(dimethylacrylamide) (PDMA), a water-soluble neutral polymer. Under conditions used for Np-SMMS of PEG with alpha-hemolysine (4M KCl, +40 mV), we obtained current blocks with polydisperse PDMA (Mn 1500 g/mol by MALDI) that were low in frequency (<0.1 Hz/ µM) short (tau <100 µs) and noisy, resulting in little mass resolution compared to PEG. We reasoned that we might take advantage of a specific salt effect of fluoride anion reported for the aHL pore3 in order to increase blocking frequency and dwell time. Using electrolyte solutions consiting of 4M K+ as cation and various proportions of Cl- and F- as anions (1:1, 2:1, 3:1, 4:1) we were able to obtain longer events (tau up to 400 µs) and higher frequencies (up to 0.15 Hz/µM), allowing significantly better mass resolution for PDMA than in 4 M KCl. However, the large noise component in the blocked current levels for PDMA as opposed to PEG remained, still compromising the peak-to-valley ratio of histograms. These findings suggest that the specific salt effect of F- on polymer-protein interaction is independent of the polymer and may be useful in tuning polymer pore interaction for a range of analytes.(1) Robertson et al. Single-Molecule Mass Spectrometry in Solution Using a Solitary Nanopore. Proc. Natl. Acad. Sci. U S A 2007, 104, 8207-8211.(2) Baaken et al. High-Resolution Size-Discrimination of Single Nonionic Synthetic Polymers with a Highly Charged Biological Nanopore. ACS Nano 2015, 9, 6443-6449.(3) Rodrigues et al. Hofmeister Effect in Confined Spaces: Halogen Ions and Single Molecule Detection. Biophys. J. 2011, 100, 2929-2935.

Rational Design of a Synthetic Peg-Like Polymer for Protein Stabilization

Organophosphorus hydrolase (OPH) is an enzyme that can deteriorate acutely toxic chemicals known as organophosphates, which are ubiquitously employed as pesticides and are thus considerable environmental hazards. However, the proclivity of OPH to aggregate in aqueous media severely hinders its biological activity. Inspired by natural chaperones, we have designed and synthesized heteropolymers to encapsulate and stabilize OPH. When OPH is complexed with the designed heteropolymer, it exhibits a drastic increase in biological activity in a buffer solution - approximately 15 times greater than the native biological activity of pure OPH in the same environment. We have confirmed via dynamic light scattering that while pure OPH aggregates in the buffer solution, the heteropolymer prevents aggregation from occurring.Two model proteins, bovine serum albumin and lysozyme, have also been studied with the heteropolymer to aid our understanding of the protein-polymer interactions that occur in our system. The structure and particle size of the different protein-polymer mixtures at several temperatures have been probed via dynamic light scattering, circular dichroism, and Fourier transform infrared spectroscopy. These characterization techniques have elucidated the mechanisms of interactions between the polymer and proteins.

Micelles from HOOC-PnBA-b-PAA-C12H15 Diblock Amphiphilic Polyelectrolytes as Protein Nanocarriers.

We investigate the potential of self-assembled nanostructures of the PnBA-b-PAA amphiphilic diblock polyelectrolyte as candidates for protein nanocarriers. Three PnBA-b-PAA copolymers with different molecular weights and PnBA/PAA weight ratios are tested. The system with the most well-defined core-shell micellar structure is chosen for complexation with lysozyme. Its solutions are found to contain well-defined core-shell micelles that are stable upon increase in solution salt content to physiological levels. Upon mixing with lysozyme we find that the protein globules accumulate preferably at the outer parts of the hydrated corona of the micelles. Increasing the protein concentration, intermicellar aggregation is enhanced in a controllable way. At high salt content the number of proteins per micelle is lower compared with the low salt content, which points to an interaction of predominantly electrostatic nature. While light scattering is very sensitive to complexation, small-angle neutron scattering is able to distinguish between the contributions from individual micelles and aggregates. This work demonstrates the use of scattering techniques to characterize protein-polymer interactions in multiple hierarchical levels.

Single-Molecule Kinetics of Protein Adsorption on Thin Nylon-6,6 Films.

Understanding and controlling protein adsorption on surfaces is critical to a range of biological and materials applications. Kinetic details that provide the equilibrium and nonequilibrium mechanisms are difficult to acquire. In this work, single-molecule fluorescence microscopy was used to study the adsorption of Alexa 555 labeled α-lactalbumin (α-LA) on two chemically identical but morphologically different polymer surfaces: flat and porous nylon-6,6 thin films. The adsorption kinetics of spatially resolved single molecule α-LA binding to nylon films were quantified by a monolayer adsorption model. The surface morphology of the porous nylon-6,6 films increased the number of adsorption sites but decreased the binding affinity compared to the flat films. Such single-molecule based kinetic studies may be extended to various protein-polymer interactions.

Best architecture of protein crystal. Database of protein-polymer interactions showing a unique role of PEG in protein crystallization

Best architecture of protein crystal. Database of protein-polymer interactions showing a unique role of PEG in protein crystallization

Bioconjugation and stabilisation of biomolecules in biosensors.

Suitable bioconjugation strategies and stabilisation of biomolecules on electrodes is essential for the development of novel and commercially viable biosensors. In the present review, the functional groups that comprise the selectable targets for practical bioconjugation methods are discussed. We focus on describing the most common immobilisation techniques used in biosensor construction, which are classified into irreversible and reversible methods. Concerning the stability of proteins, the two main types of stability may be defined as (i) storage or shelf stability, and (ii) operational stability. Both types of stability are explained, as well as the introduction of an electrophoretic technique for predicting protein–polymer interactions. In addition, solution and dry stabilisation as well as stabilisation using the covalent immobilisation of proteins are discussed including possible factors that influence stability. Finally, the integration of nanomaterials, such as magnetic particles, with protein immobilisation is discussed in relation to protein stability studies.

Charged groups synergically enhance protein imprinting in amphoteric polyacrylamide cryogels

ABSTRACTProteins are amphoteric biopolymers with unevenly charged exterior surfaces. Taking this point fully into account could accomplish ingenious recognition systems for the biological macromolecues. Molecularly imprinted polymers (MIPs) are good tools to study the interactions between polymeric matrices and template molecules. Here different protein imprinted cryogels were prepared. Imprinting factors (IFs) were determined with bovine serum albumin (BSA) as the template. The IF of the polymeric cryogel made from only acrylamide (AM) and N,N′‐methylenebisacrylamide (BisAM) is about 1.38. The introduction of charged monomers, either acrylic acid or diallylamine, would increase IFs obviously. One of the basic cryogels gave the maximum IF (about 2.0) of that type. As both acrylic acid and diallylamine were involved, IFs were further increased. An amphoteric cryogel with a suitable acid‐base ratio gave a high IF of about 3.7. Whatever used alone or both, too many added acidic or basic monomers resulted in IF reduction. Taking full advantage of charged groups in MIPs could be a good way to manipulate protein–polymer interactions. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43851.

Hot Melt Extrusion for Sustained Protein Release: Matrix Erosion and In Vitro Release of PLGA-Based Implants

The design of biodegradable implants for sustained release of proteins is a complex challenge optimizing protein polymer interaction in combination with a mini-scale process which is predictive for production. The process of hot melt extrusion (HME) was therefore conducted on 5- and 9-mm mini-scale twin screw extruders. Poly(lactic-co-glycolic acid) (PLGA) implants were characterized for their erosion properties and the in vitro release of the embedded protein (bovine serum albumin, BSA). The release of acidic monomers as well as other parameters (pH value, mass loss) during 16weeks indicated a delayed onset of matrix erosion in week 3. BSA-loaded implants released 17.0% glycolic and 5.9% lactic acid after a 2-week lag time. Following a low burst release (3.7% BSA), sustained protein release started in week 4. Storage under stress conditions (30°C, 75% rH) revealed a shift of erosion onset of 1week (BSA-loaded implants: 26.9% glycolic and 9.3% lactic acid). Coherent with the changed erosion profiles, an influence on the protein release was observed. Confocal laser scanning and Raman microscopy showed a homogenous protein distribution throughout the matrix after extrusion and during release studies. Raman spectra indicated a conformational change of the protein structure which could be one reason for incomplete protein release. The study underlined the suitability of the HME process to obtain a solid dispersion of protein inside a polymeric matrix providing sustained protein release. However, the incomplete protein release and the impact by storage conditions require thorough characterization and understanding of erosion and release mechanisms.

Protein diffusion characteristics in the hydrogels of poly(ethylene glycol) and zwitterionic poly(sulfobetaine methacrylate) (pSBMA)

Nonfouling materials such as neutral poly(ethylene glycol) (PEG) and zwitterionic poly(sulfobetaine methacrylate) (pSBMA) are ideal biocompatible materials for drug, especially protein drug delivery. The interaction behavior of protein between the nonfouling materials could cause great impact on their future applications, such as controlled release drug delivery systems. In this work, we investigated the diffusion behavior of the fluorescence-labeled model proteins (bovine serum albumin (BSA) and lysozyme (LYZ)) in nonfouling PEG, pSBMA and mixed PEG-SBMA hydrogels (SBMA:PEG 4:1, SBMA:PEG 1:4). It was observed that these four hydrogels showed varied diffusion behavior for either negatively charged BSA or positively charged LYZ due to protein-polymer interaction and the free water content in hydrogel matrix. The relatively stronger interaction between protein-PEG than protein-pSBMA could increase protein loading efficiency and control release rate by changing ratio of PEG to SBMA in the hybrid hydrogel. Moreover, it is further demonstrated the free water (freezable water) content in low cross-linked hydrogel, not the equilibrium water content (EWC), is a more accurate parameter to reflect the diffusion behavior of protein molecules. Thus, these results together provide new insights of the interactions between protein molecules and nonfouling polymers as well as the bio applications of the nonfouling polymeric hydrogels. Statement of SignificanceThis work shows that the relative stronger interaction between protein-PEG than protein-pSBMA could increase protein loading efficiency and control release rate by the change ratio of PEG to SBMA in the hydrogel, while the free water (freezable water) content in low cross-linked hydrogel, not the equilibrium water content (EWC), is a more accurate parameter to reflect the diffusion behavior of protein molecules. The impact of this work (i) gains some new insights of the interactions between protein molecules and nonfouling polymer matrixes for protein drug delivery; (ii) prompts to apply the weak PEG-protein interactions to protein drug loading and release; (iii) provides a new fundamental understanding of free water in hydrogel for protein diffusion.

Folding Behaviors of Protein (Lysozyme) Confined in Polyelectrolyte Complex Micelle.

The folding/unfolding behavior of proteins (enzymes) in confined space is important for their properties and functions, but such a behavior remains largely unexplored. In this article, we reported our finding that lysozyme and a double hydrophilic block copolymer, methoxypoly(ethylene glycol)5K-block-poly(l-aspartic acid sodium salt)10 (mPEG(5K)-b-PLD10), can form a polyelectrolyte complex micelle with a particle size of ∼30 nm, as verified by dynamic light scattering and transmission electron microscopy. The unfolding and refolding behaviors of lysozyme molecules in the presence of the copolymer were studied by microcalorimetry and circular dichroism spectroscopy. Upon complex formation with mPEG(5K)-b-PLD10, lysozyme changed from its initial native state to a new partially unfolded state. Compared with its native state, this copolymer-complexed new folding state of lysozyme has different secondary and tertiary structures, a decreased thermostability, and significantly altered unfolding/refolding behaviors. It was found that the native lysozyme exhibited reversible unfolding and refolding upon heating and subsequent cooling, while lysozyme in the new folding state (complexed with the oppositely charged PLD segments of the polymer) could unfold upon heating but could not refold upon subsequent cooling. By employing the heating-cooling-reheating procedure, the prevention of complex formation between lysozyme and polymer due to the salt screening effect was observed, and the resulting uncomplexed lysozyme regained its proper unfolding and refolding abilities upon heating and subsequent cooling. Besides, we also pointed out the important role the length of the PLD segment played during the formation of micelles and the monodispersity of the formed micelles. Furthermore, the lysozyme-mPEG(5K)-b-PLD10 mixtures prepared in this work were all transparent, without the formation of large aggregates or precipitates in solution as frequently observed in other protein-polyelectrolyte systems. Hence, the present protein-PEGylated poly(amino acid) mixture provides an ideal water-soluble model system to study the important role of electrostatic interaction in the complexation between proteins and polymers, leading to important new knowledge on the protein-polymer interactions. Moreover, the polyelectrolyte complex micelle formed between protein and PEGylated polymer may provide a good drug delivery vehicle for therapeutic proteins.

Protein Entrapment in Polymeric Mesh: Diffusion in Crowded Environment with Fast Process on Short Scales

The natural environment of proteins is a crowded environment as in cells, extracellular fluids, or during processing. Semidilute polymer solutions have been a source of rich structural and dynamical properties and mimic a crowded environment, but a proper understanding of protein dynamics in the crowded environment is far lagging. Such a study not only realizes protein’s natural environment in a crowded solution in the cell or during processing but also manifests the underlying protein–polymer interaction. By dispersing model globular proteins like α-lactalbumin (La) and hemoglobin (Hb), in aqueous solution of poly(ethylene oxide) (PEO) we mimic a crowded environment and use state-of-the-art neutron spin echo (NSE) and small-angle neutron scattering (SANS) techniques to observe the corresponding protein dynamics in semidilute polymer solution. NSE can access the fast diffusion process (Dfast) prior to the slow diffusion process on long times and length scales (Dγ). The protein dynamics in a crowded enviro...

Coacervation and precipitation in polysaccharide-protein systems.

Precipitation poses a consistent problem for the growing applications of biopolymer coacervation, but the relationship between the two types of phase separation is not well understood. To clarify this relationship, we studied phase separation as a function of pH and ionic strength, in three systems of proteins with anionic polysaccharides: β-lactoglobulin (BLG)/hyaluronic acid (HA); BLG/tragacanthin (TG); and monoclonal antibody (mAb)/HA. We found that coacervation and precipitation are intrinsically different phenomena, responsive to different factors, but their simultaneity (for example with changing pH) may be confused with transitions from one state to another. We propose that coacervate does not literally turn into precipitate, but rather that both coacervate and precipitate are in equilibrium with free protein and polyanion, so that dissolution of one and formation of the other can overlap in time. While protein-polyanion complexes must achieve neutrality for coacervation, precipitation only requires tight binding which leads to the expulsion of counterions and water molecules. The pH-dependence of phase separation, considered in terms of protein and polyion charge, revealed that the electrostatic magnitude of the protein's polymer-binding site ("charge patch") plays a key role in the strength of interaction. These findings were supported by the inhibition of precipitation, seen when the bulky side chains of TG impede close protein-polymer interactions.

The shape of protein–polymer conjugates in dilute solution

ABSTRACTProtein–polymer conjugation can significantly affect many different aspects of protein behavior, ranging from their solution properties to their ability to form solution and bulk nanostructured materials. An underlying fundamental question is how the molecular design affects the shape of the conjugate and, consequently, its properties. This work measures the molecular configuration of model protein–polymer conjugates in dilute solution using small‐angle neutron scattering (SANS) and uses quantitative model fitting to understand the shape of the molecules. Form factor measurements of four model bioconjugates of the red fluorescent protein mCherry and the polymers poly(N‐isopropylacrylamide), poly(hydroxypropyl acrylate), poly(oligoethylene glycol acrylate), and poly(ethylene glycol) show that these protein–polymer conjugates are well described by a recently developed scattering function for colloid–polymer conjugates that explicitly incorporates excluded volume interactions in the polymer configuration. In the regime where the protein does not exhibit strong interactions with the polymer, modeling the protein–polymer interactions using a purely repulsive Weeks–Chandler–Andersen potential also leads to a coarse‐grained depiction of the conjugate that agrees well with its scattering behavior. The coarse‐grained model can additionally be used for systems with varying protein–polymer interactions, ranging from purely repulsive to strongly attractive, which may be useful for conjugates with strong electrostatic or hydrophobic attractive interactions. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016, 54, 292–302

Interaction between linear PEGylated cationic block copolymers and human serum albumin

Interaction of proteins with synthetic polyelectrolytes is significant with respect to various biological applications. In this work, a detailed spectroscopic study of the binding between a series of PEGylated linear cationic block polymers (BCPs) and human serum albumin (HSA) at physiological pH has been carried out. Techniques like steady-state fluorescence spectroscopy, life-time measurements by time-correlated single photon counting (TCSPC) method and protein conformation analysis by circular dichroism (CD) measurements were employed at different temperatures. Analysis of the quenching data indicated a static quenching mechanism occurring during polymer–protein interaction. The electrostatic interaction between cationic tertiary ammonium groups of the BCPs and negatively charged amino acid residues present in HSA is the predominant force contributing to the binding process. On the other hand, the presence of PEG units in the BCPs significantly influenced the overall binding process. Conformational analysis of HSA using CD studies suggested that the linear PEG units resisted changes in the secondary structure of HSA to a significant extent, resulting in weakening of the overall binding of cationic BCPs and HSA. Detailed analysis of the thermodynamic parameters like∆G0, ∆H0and ∆S0 indicated that the binding process was entropy driven spontaneous in nature.

Non-biofouling property of well-defined concentrated polymer brushes

The non-biofouling properties of polymer brushes of poly(2-hydroxyethyl methacrylate) (PHEMA), poly(2-hydroxyethyl acrylate) (PHEA), and poly(poly(ethylene glycol) methyl ether methacrylate) (PPEGMA) were comprehensively studied by varying graft densities (i.e., semi-dilute and concentrated regimes) and the thicknesses at the dry state of 2 and 10nm. Semi-dilute polymer brushes (SDPBs) were prepared by grafting-to method and concentrated polymer brushes (CPBs) were prepared by surface-initiated atom transfer radical polymerization (SI-ATRP). The adsorptions of proteins with different sizes were investigated on the brushes by quartz crystal microbalance (QCM) from a view point of size-exclusion effect specific to the CPBs. We confirmed that due to the size exclusion effect, the CPBs of all the three much suppressed proteins adsorption and human umbilical vein endothelial cell (HUVEC) adhesion compared with the corresponding SDPBs. In order to investigate what type of proteins adsorbed on the brushes to trigger cell adhesion, we identified adsorbed proteins from fetal bovine serum on the brushes using a high-performance liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). Proteins were only detected on the SDPBs. Interestingly, the number and type of identified proteins were different on the SDPBs, indicating that chemical composition of the SDPBs affects protein adsorption, hence the cell adhesion. The adsorption mechanism on the SDPBs could be due to the combination of protein–polymer interaction and physical inclusion, whereas CPBs exhibit size exclusion effect combined with neutral hydrophilic nature of polymer, thereby, that provides excellent non-biofouling property.

A simple model for DNA bridging proteins and bacterial or human genomes: bridging-induced attraction and genome compaction

We present computer simulations of the phase behaviour of an ensemble of proteins interacting with a polymer, mimicking non-specific binding to a piece of bacterial DNA or eukaryotic chromatin. The proteins can simultaneously bind to the polymer in two or more places to create protein bridges. Despite the lack of any explicit interaction between the proteins or between DNA segments, our simulations confirm previous results showing that when the protein-polymer interaction is sufficiently strong, the proteins come together to form clusters. Furthermore, a sufficiently large concentration of bridging proteins leads to the compaction of the swollen polymer into a globular phase. Here we characterise both the formation of protein clusters and the polymer collapse as a function of protein concentration, protein-polymer affinity and fibre flexibility.

Fundamentals of nanoscale polymer-protein interactions and potential contributions to solid-state nanobioarrays.

Protein adsorption onto polymer surfaces is a very complex, ubiquitous, and integrated process, impacting essential areas of food processing and packaging, health devices, diagnostic tools, and medical products. The nature of protein–surface interactions is becoming much more complicated with continuous efforts toward miniaturization, especially for the development of highly compact protein detection and diagnostic devices. A large body of literature reports on protein adsorption from the perspective of ensemble-averaged behavior on macroscopic, chemically homogeneous, polymeric surfaces. However, protein–surface interactions governing the nanoscale size regime may not be effectively inferred from their macroscopic and microscopic characteristics. Recently, research efforts have been made to produce periodically arranged, nanoscopic protein patterns on diblock copolymer surfaces solely through self-assembly. Intriguing protein adsorption phenomena are directly probed on the individual biomolecule level for a fundamental understanding of protein adsorption on nanoscale surfaces exhibiting varying degrees of chemical heterogeneity. Insight gained from protein assembly on diblock copolymers can be effectively used to control the surface density, conformation, orientation, and biofunctionality of prebound proteins in highly miniaturized applications, now approaching the nanoscale. This feature article will highlight recent experimental and theoretical advances made on these fronts while focusing on single-biomolecule-level investigations of protein adsorption behavior combined with surface chemical heterogeneity on the length scale commensurate with a single protein. This article will also address advantages and challenges of the self-assembly-driven patterning technology used to produce protein nanoarrays and its implications for ultrahigh density, functional, and quantifiable protein detection in a highly miniaturized format.

Required polymer lengths per precipitated protein molecule in protein-polymer interaction

Protein precipitation using non-charged and charged polymers is a common method for protein purification, gaining broader interest among manufacturers in downstream processing. While during polymer- surface interactions, the formation of loops, tails and trains has been known for quite a long time, details of polymer conformation and chain length, interacting with the protein during protein precipitation are not fully discovered. Our research presents a more profound understanding of polymer-protein interaction, combining fluorescence and infrared spectroscopic measurements of proteins and polymer standards with well defined chain length to confirm different models of protein-polymer interaction. Lysozyme, chymotrypsinogen A, myoglobin and a monoclonal antibody, all of different molecular weight, isoelectric point and charge distribution at the protein surface, were used for protein-polymer precipitation. The use of polymers of various charge density and chain length showed that the required polymer length per precipitated protein (Ldef) is up to 25-times larger than the diameter of the corresponding protein, depending on the surface charge distribution of the protein, and its isoelectric point, as well as the charge density of the polymer. Our results support proposed mechanisms of polymer wrapping and loop formation for optimal charge neutralization during complexation and imply interaction of several polymer chains per precipitated protein molecule. Electrophoretic light scattering showed a qualitative correlation of the zeta potential of analyzed polymers with their corresponding Ldef values. Comparing protein precipitation behavior of long and short polymer chains, the latter exhibited reduced precipitation efficiency, visible as elevated Ldef.

Effect of polymer chemistry on globular protein–polymer block copolymer self-assembly

Changing polymer chemistry in protein–polymer conjugate block copolymers results in the formation of previously unobserved cubic phases and changes in protein–polymer interactions that create large shifts in phase transitions, providing a powerful tool for nanostructure control.

Binding characteristics between polyethylene glycol (PEG) and proteins in aqueous solution.

Polymer-protein interactions are crucial for determining the activity of both polymer and protein for many bio-related applications. Poly(ethylene glycol) (PEG) as a well-known antifouling material is often coated on surfaces to form highly solvated brushes, which exhibit excellent protein-repellent properties. However, unlike surface-induced antifouling effects, little is known about the intrinsic PEG-protein interactions in aqueous solution, which is an important yet neglected problem. Here, we investigate the interactions between PEG and proteins in aqueous solution using fluorescence spectroscopy, atomic force microscopy (AFM), and nuclear magnetic resonance (NMR). Two important characteristics, molecular weight of PEG and mass ratio of PEG : protein, are examined to determine the effect of each on PEG-protein interactions as well as binding characteristics between PEG and proteins. In contrast to too long and too short PEG chains, collective results have shown that PEG with optimal molecular weight (MW) is more capable of interacting with proteins, which induces the conformational change of proteins through more stable binding sites and stronger interactions with long chain PEG. Enhanced PEG-protein interactions are likely due to the change of hydrophilicity to amphiphilicity of PEG with increasing MWPEG. In contrast to almost none or weak interactions of PEG surfaces with proteins, this work provides new evidence to demonstrate the existence of interactions between PEG and proteins in aqueous solution, which is important not only for better understanding of the structure-activity relationship of PEG both in solution and on surfaces, but also for the rational design of new PEG-based materials for specific applications.