Levine L, Fujiki H, Yamada K, Ojika M, Gjika H B, Van Vunakis H
Department of Biochemistry, Brandeis University, Waltham, MA 02254.
Toxicon. 1988;26(12):1123-8. doi: 10.1016/0041-0101(88)90296-6.
An okadaic acid immunogen, prepared by conjugation of okadaic acid to bovine albumin with carbodiimide, was used to immunize two rabbits. The rabbits responded by producing antibodies that neutralized okadaic acid's stimulation of arachidonic acid metabolism and this neutralization increased during the course of immunization. The immune sera bound 3H-okadaic acid and this binding also increased with repeated immunization. After absorption of the rabbit IgG with a goat anti-rabbit IgG, binding was reduced greater than 99%. The binding of okadaic acid to the antibodies in one antiserum was inhibited by as little as 0.2 pmoles of unlabelled okadaic acid. The apparent association constant for binding with this antiserum was 4.17 x 10(9) M-1 (35 degrees C). Maitotoxin, teleocidin, 12-O-tetradecanoylphorbol-13-acetate, aplysiatoxin, palytoxin and brevetoxin B when tested at 29, 228, 168, 169, 3.7 and 112 pmole levels, respectively, did not inhibit binding. The serologic and biological activities of okadaic acid after incubation for 60 min in 0.01 N HCl at 35 degrees C or at 100 degrees C at pH 7.2 were unaffected.
一种通过用碳二亚胺将冈田酸与牛血清白蛋白偶联制备的冈田酸免疫原，用于免疫两只兔子。兔子产生了能中和冈田酸对花生四烯酸代谢刺激作用的抗体，并且这种中和作用在免疫过程中增强。免疫血清能结合³H-冈田酸，且随着重复免疫，这种结合也增强。用山羊抗兔IgG吸收兔IgG后，结合减少超过99%。仅0.2皮摩尔未标记的冈田酸就能抑制一种抗血清中抗体与冈田酸的结合。与该抗血清结合的表观缔合常数为4.17×10⁹ M⁻¹（35℃）。分别在29、228、168、169、3.7和112皮摩尔水平测试时， maitotoxin、teleocidin、12-O-十四烷酰佛波醇-13-乙酸酯、海兔毒素、岩沙海葵毒素和短裸甲藻毒素B均不抑制结合。在35℃于0.01 N HCl中孵育60分钟或在pH 7.2的100℃孵育后，冈田酸的血清学和生物学活性未受影响。
