An ELISA for the detection of anti-acetylcholine receptor antibodies using biotinylated alpha-bungarotoxin.
An antibody-capture enzyme immunoassay has been developed for the detection of anti-acetylcholine receptor (AChR) antibodies in tissue culture supernatants using biotinylated alpha-bungarotoxin (B alpha BGT). Immunoglobulins in culture supernatants were bound indirectly to microtitre plates via an anti-globulin antibody already coupled to polyvinyl plates. Anti-AChR antibodies were then detected by incubation with AChR crude extract. Bound AChR was revealed by incubation with B alpha BGT followed by horseradish peroxidase-conjugated avidin. This assay is specific, more sensitive than the commonly used double antibody radioimmunoassay, avoids the use of radioactive material, is practical for large numbers of samples and is particularly suitable for detecting anti-AChR antibodies in tissue culture supernatants.
Quinn
A
A
Biochemistry Department, University of Bath, U.K.
Harrison
R
R
Jehanli
A M
AM
Lunt
G G
GG
Walsh
S
S
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D001710:Biotin; D002038:Bungarotoxins; D004557:Electric Organ / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007700:Kinetics; D011863:Radioimmunoassay / Q000379:methods; D011950:Receptors, Cholinergic / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D014101:Torpedo
D000906:Antibodies; D002038:Bungarotoxins; D011950:Receptors, Cholinergic; D001710:Biotin