Purification, Quantification, and Functional Analysis of Collectins

Methods Mol Biol. 2020:2132:99-109. doi: 10.1007/978-1-0716-0430-4_10.

Abstract

Native and recombinant collectins are purified by using mannan-agarose and an anti-collectin antibody column. The use of sandwich enzyme-linked immunosorbent assay (ELISA) with two antibodies against human mannan-binding lectin (MBL) enables elucidation of the collectin concentration in the blood, serum, and plasma. The collectin sugar specificity is demonstrated by determining the concentration of saccharide required to inhibit sugar binding by 50% in a saccharide-binding assay. Biological analyses including the complement-dependent hemolysis test and several other methods are used to evaluate collectin.

Keywords: C-type lectin; CL-K1; CL-L1; Collectin; Complement; ELISA; Influenza A virus; MBL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chickens
  • Chromatography, Agarose
  • Dogs
  • Enzyme-Linked Immunosorbent Assay
  • Hemagglutinin Glycoproteins, Influenza Virus / metabolism*
  • Hemolysis
  • Humans
  • Influenza A virus / drug effects*
  • Madin Darby Canine Kidney Cells
  • Mannans / metabolism
  • Mannose-Binding Lectin / blood
  • Mannose-Binding Lectin / isolation & purification*
  • Mannose-Binding Lectin / pharmacology*

Substances

  • Hemagglutinin Glycoproteins, Influenza Virus
  • MBL2 protein, human
  • Mannans
  • Mannose-Binding Lectin