A simple method to measure anti-glycolipid antibody by using complement-mediated immune lysis of fluorescent dye-trapped liposomes.
A simple, reproducible, and micro quantity method is described to measure the antibody against glycolipid antigens. The multilamellar liposomes containing carboxyfluorescein (CF), which is self-quenched at high concentration, are prepared by vortexing the dried lipid films consisting of egg lecithin, cholesterol, phosphatidic acid and Forssman glycolipid antigen. On addition of anti-glycolipid serum plus active complement, liposome lysis occurs, and trapped CF is released. The dilution of CF in the external volume abolishes the quenching, resulting in a high fluorescence signal. Experimental conditions to measure anti-glycolipid antibody is established in this paper.
D000818:Animals; D000906:Antibodies; D003165:Complement System Proteins / Q000378:metabolism*; D005452:Fluoresceins / Q000494:pharmacology; D005456:Fluorescent Dyes; D005577:Forssman Antigen; D006017:Glycolipids / Q000276:immunology*; D006168:Guinea Pigs; D006461:Hemolysis; D007158:Immunologic Techniques; D008081:Liposomes / Q000276:immunology; D010713:Phosphatidylcholines / Q000276:immunology; D011817:Rabbits; D013997:Time Factors
D000906:Antibodies; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006017:Glycolipids; D008081:Liposomes; D010713:Phosphatidylcholines; C024098:6-carboxyfluorescein; D003165:Complement System Proteins; D005577:Forssman Antigen