Synthetic peptides VH(27-68) and VH(16-68) of the myeloma immunoglobulin M603 heavy chain and their association with the natural light chain to form an antigen binding site

Biochemistry. 1987 Dec 1;26(24):7849-55. doi: 10.1021/bi00398a047.

Abstract

A 53-residue peptide corresponding to the variable region 16-68 of the heavy chain of phosphocholine binding mouse myeloma M603 protein was synthesized by a solid-phase fragment strategy. The homogeneity of the VH(16-68) peptide was confirmed by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, amino acid analysis, and mass spectrometry. Synthetic VH(16-68) associated with the M603 light chain, and about 27% of the recombination mixture bound to phosphocholine immobilized on Sepharose as compared to a 28% binding yield obtained for the recombined natural light and heavy chains under the same conditions. The binding yield for the recombinant of the light chain with previously prepared VH(27-68) fragment was about 11%. These semisynthetic antibodies VH(27-68) and VH(16-68) light chain recombinants are forerunners of structural variants designed to study the antigen binding pocket of the M603 immunoglobulin.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Antigen-Antibody Complex / immunology*
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Polyacrylamide Gel
  • Immunoglobulin Heavy Chains / immunology*
  • Immunoglobulin Light Chains / immunology*
  • Immunoglobulin Variable Region / immunology*
  • Indicators and Reagents
  • Mass Spectrometry
  • Myeloma Proteins / immunology*
  • Peptides / chemical synthesis*
  • Peptides / immunology
  • Peptides / isolation & purification

Substances

  • Antigen-Antibody Complex
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Immunoglobulin Variable Region
  • Indicators and Reagents
  • Myeloma Proteins
  • Peptides