Effect of the light chain C-terminal serine residue on disulfide bond susceptibility of human immunoglobulin G1λ.
The light chain cysteine residue that forms an interchain disulfide bond with the cysteine residue in the heavy chain in IgG1κ is the last amino acid. The cysteine residue is followed by a serine residue in IgG1λ. Effect of the serine residue on the susceptibility of disulfide bonds to reduction was investigated in the current study using a method including reduction, differential alkylation using iodoacetic acid with either natural isotopes or enriched with carbon-13, and mass spectrometry analysis. This newly developed method allowed an accurate determination of the susceptibility of disulfide bonds in IgG antibodies. The effect of the serine residue on disulfide bond susceptibility was compared using three antibodies with differences only in the light chain last amino acid, which was either a serine residue, an alanine residue or deleted. The results demonstrated that the presence of the amino acid (serine or alanine) increased the susceptibility of the inter light and heavy chain disulfide bonds to reduction. On the other hand, susceptibility of the two inter heavy chain disulfide bonds and intrachain disulfide bonds was not changed significantly.
Liu
Hongcheng
H
Process Sciences Department, Abbott Bioresearch Center, Worcester, MA 01605, USA. Hongcheng.Liu@abbott.com
Zhong
Suju
S
Chumsae
Chris
C
Radziejewski
Czeslaw
C
Hsieh
Chung-Ming
CM
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004220:Disulfides / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007146:Immunoglobulin lambda-Chains / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D012694:Serine / Q000737:chemistry*
D004220:Disulfides; D007074:Immunoglobulin G; D007146:Immunoglobulin lambda-Chains; D012694:Serine