Blood clearance in the rat of a recombinant mouse monoclonal antibody lacking the N-linked oligosaccharide side chains of the CH2 domains.
The serum half-lives of a wild-type recombinant mouse monoclonal antibody of the IgG2b isotype and a mutant antibody differing from the wild-type antibody by a single amino acid substitution introduced into the CH2 domain, the replacement of Asn 297 by Ala to delete the conserved site of heavy chain glycosylation, were determined in the rat. The biological half-life of the aglycosyl Asn 297-Ala mutant recombinant antibody (4.8 days) was significantly shorter than that of the normally glycosylated wild-type antibody (7.4 days) by enzyme immunoassay. A similar difference between the biological half-lives of 125I-labelled aglycosyl and wild-type antibodies (2.9 and 4.0 days, respectively) was determined by gamma counting. Analysis of serum samples demonstrated that both recombinant antibodies were present in the circulation predominantly as intact monomeric IgG and revealed no differences that could account for the more rapid elimination of the aglycosyl antibody. The results of this investigation indicate that the carbohydrate residues contribute only in part to the survival of IgG in vivo and suggest that the diminished half-life of the aglycosyl antibody is due to increased catabolism in the extravascular tissues.
Wawrzynczak
E J
EJ
Drug Targeting Laboratory, Section of Medicine, Institute of Cancer Research, Sutton, Surrey, U.K.
Cumber
A J
AJ
Parnell
G D
GD
Jones
P T
PT
Winter
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000187:drug effects / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006031:Glycosylation; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007132:Immunoglobulin Isotypes / Q000378:metabolism; D008297:Male; D008657:Metabolic Clearance Rate; D009154:Mutation; D009844:Oligosaccharides / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000493:pharmacokinetics*
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007132:Immunoglobulin Isotypes; D009844:Oligosaccharides; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases