The second riboflavin-binding myeloma IgGλDOT I. Biochemical and functional characterization

Giampaolo Merlini, Reimer Bruening, Robert A. Kyle, Elliott F. Osserman

Research output: Contribution to journalArticle

Abstract

A human monoclonal immunoglobulin, IgGDOT, with flavin-binding capacity has been obtained from an elderly woman with multiple myeloma who developed yellow skin and yellow hair. The case presented a remarkable similarity with that previously reported by Farhangi and Osserman [N. Engl. J. Med. 294, 177-183 (1976)]. Purified IgGDOT was bright yellow and the ligand was identified as riboflavin and its oxidation products by thin layer chromatography, proton nuclear magnetic resonance and mass spectroscopy. Competitive binding studies with different haptens demonstrated highest affinity for riboflavin, followed by flavin mononucleotide and flavin adenine dinucleotide; no significant binding was detected for several other non-flavin compounds tested. The hapten was associated with the protein in vivo, as well as with the purified antibody. Removal of the already bound riboflavin from the combining site was associated with irreversible denaturation of the monoclonal protein. By the fluorescence quenching technique it was determined that there were 0.68 available combining sites for riboflavin molecule in IgGDOT with a binding constant of 8.5 × 108/M, while FabDOT presented 0.27 available combining sites with a binding constant of 5.1 × 108/M. The fact that 1.2 and 0.81 mol riboflavin/mol protein were already bound to IgGDOT and FabDOT, respectively, is consistent with the usual hapten/antibody stoichiometry. The heavy chain subclass of IgGDOT was identified as γ2. as in the previously reported case of riboflavin-binding protein IgGGAR. However, the λ chain subclass was different and no idiotype cross-reactivity was found.

Original languageEnglish
Pages (from-to)385-394
Number of pages10
JournalMolecular Immunology
Volume27
Issue number5
DOIs
Publication statusPublished - 1990

Keywords

  • antibody activity
  • hapten
  • monoclonal immunoglobulins
  • multiple myeloma
  • riboflavinbinding proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

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