A labeling method and purely colorimetric immunoassay based on poly-DL-lysine and a pigmented copper cluster

Suz Ann Hertzler, Ryan Brown, Matt Wilkett, Russell Boyd, Mary Ellexson, Connie Chambers, Craig Plunkett, John R. Wright

Research output: Contribution to journalArticle

Abstract

A non-biological binding action between a biotinylated, alpha-linked poly-DL-lysine (PDLL) and a pigmented, red-violet cluster anion of copper and penicillamine (RVC) may be employed as an affinity labeling method. The neutral pH association constant of this interaction is on the order of 105 M-1, and the complex survives zone electrophoresis for more than 40 min in an acetate buffer (pH 5), migrating as a cation and dissociating very slowly. Using spectrophotometric measurements at 520 nm, the RVC/PDLL binding interaction is the basis of a purely colorimetric immunoaffinity assay with a sensitivity threshold of approximately 0.3 nmol (as antibody or antigen). The method, which is based on differential elution rates, could be scaled down for capillary methods with increased sensitivity. This labeling application uses PDLL in the 30 000-70 000 g/mol molecular weight range. (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)21-25
Number of pages5
JournalMicrochemical Journal
Volume64
Issue number1
DOIs
StatePublished - 1 Jan 2000

Keywords

  • Affinity chromatography
  • Colorimetric immunoassay
  • Copper cluster
  • Poly-DL-lysine

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