Converting weak binders into infinite binders

Todd M. Corneillie, Paul A. Whetstone, Kelvin C. Lee, Jeremy P. Wong, Claude F. Meares

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Monoclonal antibody 2D12.5 binds DOTA chelates of all the rare earths with Kd ≈ 10-8 M, making it useful for the capture of probe molecules with a variety of properties. To make 2D12.5 even more useful for biological applications, we have engineered a single cysteine residue at position 54 of the heavy chain, a site proximal to the protein's binding site, so that weakly electrophilic metal complexes of (S)-2-(4-acrylamidobenzyl)-DOTA (AABD) may bind and form permanent linkages. At 37 °C, pH 7.5, all of the rare earth-AABD complexes bind permanently to the 2D12.5 G54C mutant within 5 min, in yields that correlate with their relative binding affinities. Surprisingly, indium-AABD also binds permanently in >50% yield within 5 min, despite the fact that changing the metal to indium reduces the affinity ≈100x; even copper-AABD, which has ≈10 000 x lower binding affinity than the rare earths, binds permanently in >70% yield within 2 h. However, acrylamido compounds with no measurable affinity do not bind permanently. The important practical implication is that the G54C mutant of 2D12.5 may be used for applications that include not only the rare earths, but also an unexpected range of other elements as well. This infinite binding system can exhibit selective and permanent attachment with a remarkable range of structurally related ligands, albeit at slower rates as affinities decrease.

Original languageEnglish (US)
Pages (from-to)1389-1391
Number of pages3
JournalBioconjugate Chemistry
Volume15
Issue number6
DOIs
StatePublished - Nov 2004

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

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    Corneillie, T. M., Whetstone, P. A., Lee, K. C., Wong, J. P., & Meares, C. F. (2004). Converting weak binders into infinite binders. Bioconjugate Chemistry, 15(6), 1389-1391. https://doi.org/10.1021/bc049825e