Enzyme immunometric assay for leukotriene C4.
An enzyme immunometric assay of LTC4 named SPIE-IA is described. The assay involves different sequential steps: (1) immunocapture of LTC4 by monoclonal anti-LTC4 antibodies coated on 96-well microtiter plates; (2) cross-linking of LTC4 via its amino group to the wells using glutaraldehyde; (3) treatment with HCl; (4) measurement of linked LTC4 using the same monoclonal anti-LTC4 antibodies labeled with acetylcholinesterase. A minimal detectable concentration of 2 pg/ml after 60 min of enzymatic reaction was obtained. Cross-reactivity was less than 15% with LTD4 or LTE4. The coefficient of variation was less than 6% in the 20-1000 pg/ml range. Good correlation was observed between SPIE-IA and a competitive enzyme immunoassay for biological samples. The different sequential steps of the assay are investigated.
Volland
H
H
SPI-BIO, Commissariat à l'Energie Atomique, Gif sur Yvette, France.
Vulliez Le Normand
B
B
Mamas
S
S
Grassi
J
J
Créminon
C
C
Ezan
E
E
Pradelles
P
P
D000110:Acetylcholinesterase; D000911:Antibodies, Monoclonal; D001792:Blood Platelets / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D006241:Haptens / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D017572:Leukotriene A4 / Q000097:blood; D017997:Leukotriene C4 / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D006241:Haptens; D017572:Leukotriene A4; D017997:Leukotriene C4; D000110:Acetylcholinesterase