Production of a bifunctional hybrid molecule B72.3/metallothionein-1 by protein engineering

Immunology. 1993 Apr;78(4):574-81.

Abstract

A hybrid anti-tumour B72.3 antibody/metallothionein protein B72.3MT-1 was produced by the construction of the expression vector mpSV2neo-EP1-B72.3MT-1. This vector contained the neo gene as a selection marker, the murine immunoglobulin promoter and enhancer, and the hybrid B72.3 heavy chain gene fragment with mouse metallothionein-1 cDNA gene ligated into its CH2 domain. The expression vector was transfected to the heavy chain loss mutant B72.3Mut(K) cell line. The hybrid protein B72.3MT-1 was purified from transfectant supernates using a Protein G column. We showed that the hybrid protein retained the binding reactivity for the TAG72 antigen as the original B72.3 antibody, and the metal-binding capacity of the native metallothionein molecule. Therefore, the bifunctional hybrid protein B72.3MT-1 may be very useful in cancer imaging when labelled with radionuclides such as 99mTc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / immunology
  • Antibodies, Neoplasm / biosynthesis*
  • Antibody Specificity
  • Base Sequence
  • Cell Line
  • Colonic Neoplasms / immunology
  • DNA / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Genetic Vectors
  • Humans
  • Immunoenzyme Techniques
  • Metallothionein* / genetics
  • Molecular Sequence Data
  • Protein Engineering / methods*
  • Protein Multimerization

Substances

  • Antibodies, Neoplasm
  • DNA
  • Metallothionein