A hemolytic assay for the estimation of functional mannose-binding lectin levels in human serum

Research output: Contribution to journalArticle

Abstract

A simple assay was developed to estimate functional mannose-binding lectin (MBL) levels in serum based on the principle of yeast-induced bystander lysis of chicken erythrocytes (ChE). The assay is sensitive to inhibition by ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) (which allows alternative pathway activation), ethylene diamine tetraacetic acid (EDTA), mannose, N-acetylglucosamine and C1 esterase inhibitor (C1-INH), whereas it was not inhibited by galactose. A high-titer human anti-mannan antibody-containing serum with 0.06 mug MBL/ml gave a functional signal corresponding to 0.12 mug equivalents MBL/ml, indicating that anti-mannan antibodies are poorly hemolytic in the assay. The assay is well suited for the large-scale testing of patient samples for a functional MBL pathway of complement activation.

Details

Authors
  • S Kuipers
  • PC Aerts
  • Anders Sjöholm
  • T Harmsen
  • H van Dijk
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Immunology in the medical area

Keywords

  • bystander hemolysis, functional assay, MBL, lectin pathway, Saccharomyces cerevisiae
Original languageEnglish
Pages (from-to)149-157
JournalJournal of Immunological Methods
Volume268
Issue number2
Publication statusPublished - 2002
Publication categoryResearch
Peer-reviewedYes