Measurement of myelin basic protein and of anti-basic protein antibodies by ELISA utilizing biotinylated antibodies.
Immunoglobulins were conjugated to peroxidase by the biotin-avidin method and used in ELISA systems for measuring myelin basic protein (MBP) and anti-MBP antibodies. To measure concentration of MBP, microplate wells were coated with affinity purified rabbit anti-MBP antibodies and incubated with varying concentrations of MBP. Bound antigen was measured by incubating with biotinylated anti-MBP antibodies and avidin-peroxidase. As little as 0.2 ng/ml of MBP could be measured by this assay. To measure anti-MBP antibodies, microplate wells were coated with human MBP and incubated with varying concentrations of affinity purified rabbit anti-human MBP antibodies. Binding was measured by incubating with either peroxidase-conjugated anti-rabbit antibodies or biotinylated anti-rabbit antibodies and avidin peroxidase. The two methods were equally sensitive. The avidin-biotin method for enzyme conjugation promises to be a useful and versatile tool for ELISA systems.
D000818:Animals; D001323:Autoantibodies / Q000032:analysis*; D001360:Avidin / Q000378:metabolism; D001710:Biotin / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D004676:Myelin Basic Protein / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D013388:Succinimides
D001323:Autoantibodies; D004676:Myelin Basic Protein; D013388:Succinimides; C033788:biotinyl N-hydroxysuccinimide ester; D001360:Avidin; D001710:Biotin