Protein hydrophilic copolymers

Since the hydrophobicity of styrene- or alkyl methacrylate-based monolithic matrices is too high to make them useful for hydrophobic interaction chromatography, porous monoliths based on highly hydrophilic copolymers of acrylamide and methylenebisacrylamide were developed [70,135]. The hydrophobicity of the matrix required for the successful separations of proteins is controlled by the addition of butyl methacrylate to the polymerization mixture. The suitability of this rigid hydrophilic monolith for the separation of protein mixtures is demonstrated in Fig. 21, which shows the rapid separation of five proteins in less than 3 min using a steeply decreasing concentration gradient of ammonium sulfate. 

Lens hazing and protein deposition are common problems for wearers of soft contact lenses. Previous experiments with hydrophobic-hydrophilic copolymers exposed to plasma showed protein adsorption to be minimal at intermediate copolymer compositions. Adsorption of proteins from artificial tear solutions to a series of polymers and copolymers ranging in composition from 100% poly (methyl methacrylate) (PMMA) to 100% poly(2-hydroxyethyl methacrylate) (PH EM A) was measured. The total protein adsorption due to the three major proteins in tear fluid (lysozyme, albumin, and immunoglobulins) was at a minimum value at copolymer compositions containing 50% or less PH EM A. The elution of the adsorbed proteins from these polymers and copolymers with various solutions also was investigated to assess the binding mechanism. 

The accumulation of proteins on contact lenses has long been viewed as an undesirable event. In this study, the effect of polymer composition on both the total amount of protein on the materials, and on the specific proteins on each polymer composition was documented. The importance of these factors for biological response is not known, so this situation remains a fertile area for investigation. This study also demonstrated that a linear variation in material composition will not necessarily result in a linear variation in absorbed layer protein composition. The minima and maxima noted at intermediate copolymer compositions have strong implications for both understanding the mechanism of protein adsorption and for biological response. Investigation is underway to explore further protein interaction with hydrophobic-hydrophilic copolymer materials. 

Micellar nanocarriers have already been applied successfully for delivery of hydro-phobic drugs [86]. These carriers are usually the product of self-assembled block copolymers, consisting of a hydrophilic block and a hydrophobic block. Generally, an ELP with a transition temperature below body temperature is used as hydrophobic block and the hydrophilic block can be an ELP with a transition temperature above body temperature or another peptide or protein. The EPR effect also directs these types of carriers towards tumor tissue. 

Many kinds of nonbiodegradable vinyl-type hydrophilic polymers were also used in combination with aliphatic polyesters to prepare amphiphilic block copolymers. Two typical examples of the vinyl-polymers used are poly(/V-isopropylacrylamide) (PNIPAAm) [149-152] and poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) [153]. PNIPAAm is well known as a temperature-responsive polymer and has been used in biomedicine to provide smart materials. Temperature-responsive nanoparticles or polymer micelles could be prepared using PNIPAAm-6-PLA block copolymers [149-152]. PMPC is also a well-known biocompatible polymer that suppresses protein adsorption and platelet adhesion, and has been used as the hydrophilic outer shell of polymer micelles consisting of a block copolymer of PMPC -co-PLA [153]. Many other vinyl-type polymers used for PLA-based amphiphilic block copolymers were also introduced in a recent review [16]. 

The following protocol for passive adsorption is based on methods reported for use with hydrophobic polymeric particles, such as polystyrene latex beads or copolymers of the same. Other polymer particle types also may be used in this process, provided they have the necessary hydrophobic character to promote adsorption. For particular proteins, conditions may need to be optimized to take into consideration maximal protein stability and activity after adsorption. Some proteins may undergo extensive denaturation after immobilization onto hydrophobic surfaces therefore, covalent methods of coupling onto more hydrophilic particle surfaces may be a better choice for maintaining native protein structure and long-term stability. 

This strategy was first realized by Lozinsky et al., who studied the redox-initiated free-radical copolymerization of thermosensitive N-vinylcaprolactam with hydrophilic N-vinylimidazole at different temperatures, as well as by Chi Wu and coworkers. Lozinsky presents an extensive review of the experimental approaches, both already described in the literature and potential new ones, to chemical synthesis of protein-like copolymers capable of forming core-shell nanostructures in a solution. 

Abstract Protein-like copolymers were first predicted by computer-aided biomimetic design. These copolymers consist of comonomer units of differing hydrophilicity/hydro-phobicity. Heterogeneous blockiness, inherent in such copolymers, promotes chain folding with the formation of specific spatial packing a dense core consisting of hydrophobic units and a polar shell formed by hydrophilic units. This review discusses the approaches, those that have already been described and potential approaches to the chemical synthesis of protein-like copolymers. These approaches are based on the use of macromolecular precursors as well as the appropriate monomers. In addition, some specific physicochemical properties of protein like copolymers, especially their solution behaviour in aqueous media, are considered. 

Nonetheless, one cannot exclude the probability of a successful combination of these prerequisites (as was the case with poly[(NiPAAm-co-GMA)-g-PEO considered above]) that will allow us to obtain, using the chemical colouring approach, the protein-like HP-copolymers with a dense hydrophobic core wrapped by the hydrophilic shell. Such a shell should be capable of efficiently protecting the temperature-responsive macromolecules against pronounced interchain hydrophobic interactions and precipitation at temperatures significantly higher than those at which the copolymers of the same total monomer composition—but with a non-protein-like primary sequence of comonomer units—are in the soluble state. 

A general idea related to the preparation of protein-like copolymers through the co-polymerization or co-polycondensation of the mixtures of comonomers with differing hydrophilicity/hydrophobicity has been described in Sect. 2.1. Scheme 4 demonstrates the multi-step operations used in the first successful realization [26,27] of such an approach in a free radical polymerization process. 

The transition enthalpies of the s- and p-fractions obtained from the feed with a comonomer molar ratio of 85 15 were equal to 6 and 7 J/g, respectively, i.e. the values are very close. This, therefore, can be indicative of almost the same average length of oligoNVCl blocks. Moreover, as we have already stressed, the fractions also had virtually the same final comonomer composition. However, since the solution properties of these fractions are drastically different, one can draw the conclusion that this is apparently due to a specific distribution of hydrophobic and hydrophilic residues along the polymer chains. In turn, because of all the properties that are exhibited by the s-fraction, this fraction can be considered to be a protein-like copolymer [27]. 

Corresponding approaches were developed in all the research methods theoretical, computer simulation, and, moreover, experimental. Thus, copolymers were synthesized in vitro, which form non-aggregating structures of the type hydrophobic core-hydrophilic shell. The structure of such copolymers is similar in this respect to that of protein macromolecules [125-127]. 

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