Microheterogeneity of mouse antidextran monoclonal antibodies

Electrophoresis
Y MimuraK Nakamura

Abstract

Mouse antidextran monoclonal antibodies showed microheterogeneity which was analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Not only the heavy (H) chains but also the light (L) chains were heterogeneous in terms of isoelectric point (pI). The higher the pI, the more prominent the H chain spots. To demonstrate the cause of the microheterogeneity an IgG1 monoclonal antibody (mAb 35.8.2H) was examined especially for involvement of the sugar moiety in the microheterogeneity. The glycosylated region was determined in the Fc portion from serine 239 to methionine 309 by a glycan detection method using mild periodate oxidation, which confirms that the sugar chain is attached to the conserved glycosylation site of asparagine 297. However, charge heterogeneity of the H chain was not entirely attributed to the Fc because the papain digest of the antibody was separated into two Fc spots, a few Fd spots and two L chain spots by 2-D PAGE. This indicates that factors other than the sugar moiety are responsible for charge heterogeneity of IgG monoclonal antibody. On the other hand, the H chain isoforms of lower pI were shown to be more susceptible to V8 protease by peptide mapping. This result strongly suggests the o...Continue Reading

References

Apr 1, 1985·The Journal of Experimental Medicine·S H ClarkeM Weigert
Feb 1, 1972·The Biochemical Journal·J Svasti, C Milstein

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Citations

Apr 10, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jinzhi Chen, Cheng S Lee
Nov 15, 1996·Journal of Chromatography. B, Biomedical Applications·S AnastaseJ Jozefonvicz
Oct 29, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·K Nakamura, K Takeo
Dec 17, 2009·Journal of Pharmaceutical Sciences·Hui ZhaoKurt Forrer
Sep 11, 2007·Journal of Pharmaceutical Sciences·Hongcheng LiuJoanne Sun
Jul 27, 2010·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Sascha HagemannRichard Dodel
Jul 15, 2006·Advanced Drug Delivery Reviews·Ann L Daugherty, Randall J Mrsny
Jun 16, 2017·Clinical and Experimental Immunology·Y MurataY Mimura

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