An improved method for conjugating monoclonal antibodies with N-hydroxysulfosuccinimidyl DOTA

Bioconjug Chem. 2001 Mar-Apr;12(2):320-4. doi: 10.1021/bc0000886.

Abstract

A simple, water-soluble procedure for conjugation of monoclonal antibodies to 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA) has been improved by optimizing pH, buffer, and temperature conditions for the preparation of N-hydroxysulfosuccinimidyl DOTA and its conjugation to the human/murine chimeric anti-carcinoembryonic antigen antibody cT84.66. This improved method results in a 6-fold increase in conjugation efficiency, a 3-7-fold decrease in antibody cross-linking, a more homogeneous population of conjugate species, and a 5-fold decrease in the quantities of reagents needed for conjugation. The cT84.66-DOTA conjugate was labeled to high specific activity with 111In, 90Y, 88Y, 64Cu, and 67Cu, affording near-quantitative incorporation of the majority of these radiometals. This improved conjugation procedure facilitates large-scale production and radiometal labeling of cT84.66-DOTA for clinical radioimmunotherapy trials.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Neoplasm / chemistry*
  • Buffers
  • Carcinoembryonic Antigen / immunology*
  • Chimerin Proteins / chemistry
  • Heterocyclic Compounds, 1-Ring / chemistry*
  • Humans
  • Hydrogen-Ion Concentration
  • Immunoconjugates / chemistry*
  • Isoelectric Focusing
  • Metals / chemistry
  • Mice
  • Radioimmunodetection
  • Radioisotopes / chemistry
  • Succinimides / chemistry*
  • Temperature

Substances

  • Antibodies, Monoclonal
  • Antibodies, Neoplasm
  • Buffers
  • Carcinoembryonic Antigen
  • Chimerin Proteins
  • Heterocyclic Compounds, 1-Ring
  • Immunoconjugates
  • Metals
  • Radioisotopes
  • Succinimides
  • 1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid
  • N-hydroxysulfosuccinimide