Polystyrene proteins

Ion exchange resins are also useful for demineralising biochemical preparations such as proteins. Removal of metal ions from protein solutions using polystyrene-based resins, however, may lead to protein denaturation. This difficulty may be avoided by using a weakly acidic cation exchanger such as Bio-Rex 70. 

Size exclusion was first noted in the late fifties when separations of proteins on columns packed with swollen maize starch were observed (Lindqvist and Storgards, 1955 Lathe and Ruthven, 1956). The run time was typically 48 hr. With the advent of a commercial material for size separation of molecules, a gel of cross-linked dextran, researchers were given a purposely made material for size exclusion, or gel filtration, of solutes as described in the classical work by Porath and Flodin (1959). The material, named Sephadex, was made available commercially by Pharmacia in 1959. This promoted a rapid development of the technique and it was soon applied to the separation of proteins and aqueous polymers. The work by Porath and Flodin promoted Moore (1964) to apply the technique to size separation, gel permeation chromatography of organic molecules on gels of lightly cross-linked polystyrene (i.e., Styragel). 

We ve seen on several occasions in previous chapters that a polymer, whether synthetic or biological, is a large molecule built up by repetitive bonding together of many smaller units, or monomers. Polyethylene, for instance, is a synthetic polymer made from ethylene (Section 7.10), nylon is a synthetic polyamide made from a diacid and a diamine (Section 21.9), and proteins are biological polyamides made from amino acids. Note that polymers are often drawn by indicating their repeating unit in parentheses. The repeat unit in polystyrene, for example, comes from the monomer styrene. 

The pore structure of most cross-linked polystyrene resins are the so called macro-reticular type which can be produced with almost any desired pore size, ranging from 20A to 5,000A. They exhibit strong dispersive type interaction with solvents and solutes with some polarizability arising from the aromatic nuclei in the polymer. Consequently the untreated resin is finding use as an alternative to the C8 and Cl8 reverse phase columns based on silica. Their use for the separation of peptide and proteins at both high and low pH is well established. 

Most frequently, SEC with dextran-, pullulan-, or polystyrene calibration standards has been used to characterize the molecular properties of xylans. However, as for viscometric studies [108], a sufficient solvent ionic strength is a prerequisite for useful SEC measurements of charged polysaccharides, including glucuronoxylans [111-113]. An advantage of the SEC technique is that the presence of protein and phenolic components or oxidative changes can be detected by simultaneous ultraviolet (UV) detection. 

Adamski, RP Anderson, JL, Configurational Effects on Polystyrene Rejection from Micro-porosou Membranes, Journal of Polymer Science Part B Polymer Physics 25, 765, 1987. Adler, PM, Porous Media, Geometry and Transports Butterworth-Heinemann Boston, 1992. Afeyan, NB Fulton, SP Regnier, FE, Perfusion Chromatography Packing Materials for Proteins and Peptides, Journal of Chromatography 544, 267, 1991. 

Yang, Y.-B., Harrison, K., and Kindsvater, J., Characterization of a novel stationary phase derived from a hydrophilic polystyrene-based resin for protein cation-exchange high-performance liquid chromatography, /. Chromatogr. A, 723, 1, 1996. 

Ivanov, A.R., Zang, L., Karger, B. L. (2003). Low-attomole electrospray ionization MS and MS/ MS analysis of protein tryptic digests using 20 pm-i.d. polystyrene-divinylbenzene monolithic capillary columns. Anal. Chem. 75, 5306-5316. 

Figure 3. Protein decoupled 13C NMR spectrum at 75 MHz of 17,500 MW polystyrene at 20% w/v in CDCU/CHCU (lower trace). One percent cross-linked polystyrene (Biobeads SX-1) in the same solvent (upper trace).

Kumada Y, Shiritani Y, Hamasaki K, Ohse T, Kishimoto M (2009) High biological activity of a recombinant protein immobilized onto polystyrene. Biotechnol J 4 1178-1189... 

The following protocol describes the modification of a protein with mono(lactosylamido) mono(succinimidyl)suberate. The reagent is available from Thermo Fisher. The use of this reagent to couple to amine-containing surfaces, such as polystyrene beads, also has been done using similar reaction conditions (Vetter et al., 1995). 

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. 

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