Isolation and characterization of a monomethioninesulfoxide variant of interferon alpha-2b

被引：29

作者：

Gitlin, G ^{[1
]}

Tsarbopoulos, A ^{[1
]}

Patel, ST ^{[1
]}

Sydor, W ^{[1
]}

Pramanik, BN ^{[1
]}

Jacobs, S ^{[1
]}

Westreich, L ^{[1
]}

Mittelman, S ^{[1
]}

Bausch, JN ^{[1
]}

机构：

[1] SCHERING PLOUGH CORP,RES INST,KENILWORTH,NJ 07033

来源：

PHARMACEUTICAL RESEARCH | 1996年 / 13卷 / 05期

关键词：

protein mapping; mass spectrometry; methionine oxidation;

D O I：

10.1023/A:1016059902645

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Purpose. To isolate and characterize a monomethioninesulfoxide variant of the commercially available therapeutic protein interferon alpha-2b. Methods. The methionine (Met)-oxidized variant was isolated by reverse-phase high performance liquid chromatography and characterized by SDS-PAGE, peptide mapping and mass spectrometric analysis of the trypsin/V8-generated peptide fragments. The biological and immunological activities of the isolated variant were also evaluated. Results. The rHuIFN alpha-2b variant was found to contain a Met sulfoxide residue at position ill of the rHuIFN alpha-2b molecule. The far-UV CD spectra showed a slight loss of alpha-helical content and an increase in the beta-sheet contribution. The CD spectra indicate that both chromatographic conditions and Met oxidation contribute to the observed secondary structure changes. Both interferon alpha-2b main component and its methionine-oxidized variant showed different reactivity to monoclonal antibodies employed in immunoassays for the protein. Conclusions. A monomethioninesulfoxide rHuIFN alpha-2b variant was found to be present in the rHuIFN alpha-2b bulk drug substance in solution. The Met(111) residue was identified as Met sulfoxide by comparative tryptic/V8 mapping and mass spectrometric analysis. Nevertheless, the oxidation of the Met(111) residue did not seem to have a detectable effect on the biological activity of the molecule.

引用

页码：762 / 769

页数：8

共 18 条

[1]

BECKER GW, 1988, BIOTECHNOL APPL BIOC, V10, P326

[2] MASS-SPECTROMETRIC DETERMINATION OF THE AMINO-ACID-SEQUENCE OF PEPTIDES AND PROTEINS [J].