A simple radioimmunoassay to determine binding of antibodies to lipid antigens

J Immunol Methods. 1980;38(1-2):85-93. doi: 10.1016/0022-1759(80)90333-6.

Abstract

A solid-phase radioimmunoassay to determine the binding of antibodies to lipid antigens is described. Polyvinyl chloride microtiter plates were coated with lipid antigens by placing in each well 25--100 microliters of solutions of the antigens in ethanol and evaporation of the solvent. The wells were than washed with 0.3% gelatine solution to remove loosely bound antigens and to saturate the plastic with an unrelevant protein to prevent unspecific binding of antibodies to the plates. About one-third of [3H] phosphatidylcholine adsorbed in this way (out of 1 nmole) was firmly attached to the plastic and was not washed away during a standard assay. The rest, loosely bound lipid, was washed away in the first 5 washings before the primary antibodies were applied. The technique is rapid and convenient and is as sensitive as a conventional solid-phase radioimmunoassay with protein antigens. It requires minute amounts of antigen. Less than 10(-12) moles of lipid antigen can be detected by this technique.

MeSH terms

  • Animals
  • Antigen-Antibody Reactions*
  • Antigens / immunology*
  • Goats
  • Membrane Lipids / immunology*
  • Rabbits
  • Radioimmunoassay / methods*

Substances

  • Antigens
  • Membrane Lipids