Application of the Sandwich Enzyme Immunoassay for Determination of Batroxobin in Various Animal Plasma

• Published in: YAKUGAKU ZASSHI Vol. 106; no. 4; pp. 330 - 334
• Main Authors: SUGAI, KEN-ICHI, IMAMURA, YASUSHI, MIHASHI, SUSUMU, MORI, NOBORU, BABA, SHIGEO
• Format: Journal Article
• Language: English; Japanese
• Published: Japan The Pharmaceutical Society of Japan 01-04-1986
• Subjects: Animals; batroxobin; Batroxobin - blood; defibrinogenating agent; diisopropylfluorophosphate; Dogs; enzyme immunoassay; Fibrinolytic Agents - blood; Immunoenzyme Techniques; Macaca fascicularis; Male; Mice; Peptide Hydrolases - blood; plasma; Rabbits; Rats; Species Specificity; urokinase
• ISSN: 0031-6903; 1347-5231
• DOI: 10.1248/yakushi1947.106.4_330

In order to estimate trace amounts of batroxobin, a defibrinogenating agent, we established sandwich enzyme immunoassay of this substance in the plasma sample. The Fab' fragment was labeled with horseradish peroxidase by using 4-(maleimidomethyl)-cyclohexane-1-carboxlic acid succimide ester as a coupling reagent. The conjugate thus obtained could be used to determine batroxobin concentration in the plasma sample up to 25 pg/tube (minimum detectable concentration) with the variable coefficient of 8.4%(within-run) and 13.8%(between-run). It was observed that the recovery of batroxobin from the plasma in several animal species ranges from 25% to 90%, when determined by this method. Satisfactory result was obtained by treatment of plasma with diisopropylfluorophosphate or by fibrinolysis with urokinase.