Oxidation of recombinant human interleukin-2 by potassium peroxodisulfate

被引:21
作者
Cadée, JA
van Steenbergen, MJ
Versluis, C
Heck, AJR
Underberg, WJM
den Otter, W
Jiskoot, W [1 ]
Hennink, WE
机构
[1] UIPS, Dept Pharmaceut, Utrecht, Netherlands
[2] UIPS, Dept Biomol Mass Spectrometry, Utrecht, Netherlands
[3] Bijvoet Ctr Biomol Res, Utrecht, Netherlands
[4] Univ Utrecht, UIPS, Dept Biomed Anal, Utrecht, Netherlands
[5] Univ Utrecht, Dept Biochem Cell Biol & Histol, Utrecht, Netherlands
关键词
oxidation; interleukin-2; methionine; peroxodisulfate; mass spectrometry;
D O I
10.1023/A:1012213108319
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Purpose. The oxidation of recombinant human interleukin-2 (rhIL-2) by potassium peroxodisulfate (KPS) with or without NNN',N'-tetramethylethylenediamine (TEMED), which are used for the preparation of dextran-based hydrogels, was investigated. Methods. The oxidation of (derivatives of) methionine, tryptophan, histidine and tyrosine, as well as rhIL-2 was investigated. Both the oxidation kinetics (RP-HPLC) and the nature of the oxidation products (mass spectrometry) were studied as a function of the KPS and TEMED concentration, and the presence of a competitive antioxidant, methionine. Results. Under conditions relevant for the preparation of rhIL-2 loaded hydrogels, only methionine and tryptophan derivatives were susceptible to oxidation by XPS, The oxidation of these compounds was inhibited once TEMED was present, suggesting that the peroxodisulfate anion, rather than the radicals formed in the presence of TEMED, is the oxidative species. KPS only induced oxidation of the four methionines present in rhIL-2, whereas the tryptophan residue remained unaffected. The radicals, formed after KPS decomposition by TEMED, induced some dimerization of rhIL-2. The oxidation of rhIL-2 could be substantially reduced by the addition of methionine, or by pre-incubation of KPS with TEMED. Conclusions. Only the methionine residues in rhIL-2 are oxidized by KPS. The extent of oxidation can be minimized by a proper selection of the reaction conditions.
引用
收藏
页码:1461 / 1467
页数:7
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