Monoclonal anti-histamine antibody. Preparation, characterization and application to enzyme immunoassay of histamine.
An enzyme immunoassay to measure histamine has been developed. A histamine-bovine serum albumin conjugate was prepared using 1,4-benzoquinone as the coupling agent and was employed to immunize mice for the preparation of monoclonal antibodies against histamine. After an initial screening to identify antigen-binding monoclonal antibodies the clones were isolated by limiting dilution cloning, grown in ascites and antibodies which had been secreted into the ascitic fluid were precipitated by ammonium sulphate at 50% saturation. A systematic approach for the determination of epitope specificities of monoclonal antibodies was performed. It was found that for the most specific antibody the main epitope encompassed the 2-histaminyl-1,4-benzoquinone moiety and that the KD value determined by indirect ELISA was 1.5 X 10(-8) M for the hapten part of the immunogen and 4.6 X 10(-10) M for a histamine-Bq-ovalbumin conjugate. The selected monoclonal antibody could not recognize histidine or methyl-histamine. Using this antibody, we developed an enzyme immunoassay for histamine and pg amounts could be detected. The same assay was used to quantify the allergic release of histamine from guinea pig lung mast cells. Results obtained either by the present enzyme immunoassay or by a fluorometric assay were closely correlated (correlation coefficient r = 0.9702, n = 37).
Guesdon
J L
JL
Chevrier
D
D
Mazié
J C
JC
David
B
B
Avrameas
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D016227:Benzoquinones; D000939:Epitopes; D005260:Female; D006241:Haptens / Q000276:immunology; D006632:Histamine / Q000276:immunology*; D008722:Methods; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D011809:Quinones / Q000276:immunology
D000911:Antibodies, Monoclonal; D016227:Benzoquinones; D000939:Epitopes; D006241:Haptens; D011809:Quinones; C004532:quinone; D006632:Histamine