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- W1968045576 abstract "Abstract It has been found that all commercially available carrier ampholytes (the amphoteric buffers used to create and stabilize the pH gradient in isoelectric focusing, IEF) (e.g., Ampholine, Pharmalyte) are extensively oxidized by persulphate during gel polymerization. These oxidized species are formed for all pH ranges, from acidic to basic, and exhibit chromophoric peaks in the 300–360 nm range. It is believed that these oxidized products are N-oxides (possibly even nitrosamines). Owing to their high redox potentials during IEF, especially in alkaline pH ranges, they are able to capture two H+ and two electrons and be reduced, while oxidizing in proteins SH group of Cys to —S—S— (possibly even to cysteic acid). This generates artefactual band heterogeneity, not present in the starting sample, which could never be eliminated as, on rerunning the zone, additional oxidation will take place. In such cases, the presentation of proof of sample homogeneity to agencies such as the FDA, for recombinant DNA products for human consumption, might be impossible. Such a cause of artefactual sample heterogeneity can be virtually eliminated by simply not exposing the carrier ampholytes to persulphate. In the proposed IEF protocol, “empty” gels are polymerized, washed, dried and reswollen in the desired pH interval." @default.
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- W1968045576 date "1989-01-01" @default.
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- W1968045576 title "Carrier ampholyte-mediated oxidation of proteins in isoelectric focusing" @default.
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- W1968045576 doi "https://doi.org/10.1016/s0021-9673(01)89683-7" @default.
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