Identifying amino acid residues that influence plasma clearance of murine IgG1 fragments by site-directed mutagenesis

Eur J Immunol. 1994 Mar;24(3):542-8. doi: 10.1002/eji.1830240308.

Abstract

Site-directed mutagenesis has been used to change amino acid residues of a recombinant Fc-hinge fragment derived from the murine immunoglobulin (Ig)G1 molecule, and the effects of these mutations on the pharmacokinetics of the Fc-hinge fragment have been determined. Specifically, Ile-253, His-310 and Gln-311 of the CH2 domain and His-433 and Asn-434 of the CH3 domain have been changed. In the three dimensional structure of an antibody, these amino acids are in close proximity to each other at the CH2-CH3 domain interface. The mutated Fc-hinge fragments have been purified from recombinant Escherichia coli cells and their pharmacokinetic parameters determined in mice and compared with those of the wild-type Fc-hinge fragment. The results show that the site of the IgG1 molecule that controls the catabolic rate (the 'catabolic site') is located at the CH2-CH3 domain interface and overlaps with the Staphylococcal protein A binding site.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Circular Dichroism
  • Immunoglobulin Fc Fragments / chemistry
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / metabolism*
  • Metabolic Clearance Rate
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Structure, Secondary
  • Staphylococcal Protein A / metabolism
  • Structure-Activity Relationship

Substances

  • Immunoglobulin Fc Fragments
  • Immunoglobulin G
  • Staphylococcal Protein A