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Isolation and characterization of a VH domain fragment from monoclonal rat IgG of IgG1 and IgG2a subclasses
J Rousseaux1, R Rousseaux-Prevost, H Bazin
1Unité CNRS no. 409, Institut de Recherches sur le Cancer de Lille, France.
Digestion with trypsin of monoclonal rat IgG of IgG1 and IgG2a subclasses produced two fragments, isolated only in dissociating media. The larger fragment (mol. wt. 120,000 Da) was comprised of the two light chains covalently bound to shortened gamma chains. Amino acid sequence of the shortened gamma chain indicated that the site of cleavage is located at the beginning of the C gamma 1 domain at a position homologous to residue 139 of mouse gamma heavy chain of IgG1 MOPC 21. The smaller fragment (mol. wt. 13,000 Da) was found to consist of the entire variable domain of the heavy chain and probably a short stretch of the C gamma 1 domain. The unique susceptibility of rat monoclonal IgG1 and IgG2a is likely to be the result of the presence of a lysine residue in a loop of C gamma 1 domain, which therefore is accessible to trypsin. Tryptic cleavage of rat monoclonal antibodies of IgG1 and IgG2a subclasses can be considered as a simple method to produce a fragment related to the VH domain.
Digestion with trypsin of monoclonal rat IgG of IgG1 and IgG2a subclasses produced two fragments, isolated only in dissociating media. The larger fragment (mol. wt. 120,000 Da) was comprised of the two light chains covalently bound to shortened gamma chains. Amino acid sequence of the shortened gamma chain indicated that the site of cleavage is located at the beginning of the C gamma 1 domain at a position homologous to residue 139 of mouse gamma heavy chain of IgG1 MOPC 21. The smaller fragment (mol. wt. 13,000 Da) was found to consist of the entire variable domain of the heavy chain and probably a short stretch of the C gamma 1 domain. The unique susceptibility of rat monoclonal IgG1 and IgG2a is likely to be the result of the presence of a lysine residue in a loop of C gamma 1 domain, which therefore is accessible to trypsin. Tryptic cleavage of rat monoclonal antibodies of IgG1 and IgG2a subclasses can be considered as a simple method to produce a fragment related to the VH domain.