Site-specific modification of a single-chain antibody using a novel glyoxylyl-based labeling reagent

Zhan G. Zhao, Jin S. Im, Kit Lam, Douglas F. Lake

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

A novel, highly specific protein modification approach is described. By using conventional molecular cloning techniques, a protein can be constructed and expressed such that the N-terminal residue is replaced by cysteine. Its 1,2-aminothiol structure reacts very specifically with a glyoxylyl group at pH 7 or below, forming a relatively stable thiazolidine bridge. Therefore, a glyoxylyl-based labeling agent (e.g., radioactive tags, fluorescent probes, biotin) can be used to specifically modify a protein at its N-terminus. To highlight this novel approach, a recombinant anti-insulin single chain antibody (scFv) was specifically biotinylated at its N-terminus even in the presence of other proteins in the total cell lysate. The glyoxylyl-biotinylated scFv retained binding activity similar to unmodified scFv.

Original languageEnglish (US)
Pages (from-to)424-430
Number of pages7
JournalBioconjugate Chemistry
Volume10
Issue number3
DOIs
StatePublished - May 1999
Externally publishedYes

Fingerprint

Single-Chain Antibodies
Antibodies
Labeling
Proteins
Thiazolidines
Insulin Antibodies
Cloning
Insulin
Molecular Cloning
Biotin
Fluorescent Dyes
Cysteine

ASJC Scopus subject areas

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

Cite this

Site-specific modification of a single-chain antibody using a novel glyoxylyl-based labeling reagent. / Zhao, Zhan G.; Im, Jin S.; Lam, Kit; Lake, Douglas F.

In: Bioconjugate Chemistry, Vol. 10, No. 3, 05.1999, p. 424-430.

Research output: Contribution to journalArticle

Zhao, Zhan G. ; Im, Jin S. ; Lam, Kit ; Lake, Douglas F. / Site-specific modification of a single-chain antibody using a novel glyoxylyl-based labeling reagent. In: Bioconjugate Chemistry. 1999 ; Vol. 10, No. 3. pp. 424-430.
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