Yamamoto I, Morishita K
J Immunoassay. 1986;7(1-2):17-35. doi: 10.1080/01971528608063044.
A sensitive and specific enzyme immunoassay for an immunomodulator PLATONIN (NK 19) has been developed and applied to the evaluation of pharmacokinetics in human and rats. NK 19 analog, succinyl NK 19 has been synthesized and coupled to bovine serum albumin. This conjugate is immunogenic and when injected into rabbits antibodies of high affinity for NK 19 were obtained. Some light decomposed products of Platonin were of lower cross-reactivity with these antibodies. For competitive reactions, these antibodies were incubated with a mixture of succinyl NK 2900-beta-D-galactosidase conjugate and standard NK 19 or sample. Then, antibody-bound enzyme-hapten were separated from free one using anti-rabbit IgG immobilized on polystyrene balls. Activity of the enzyme on the solid phase was fluorometrically determined. The present immunoassay allows detection of as low as 10 pg of NK 19 in the biological fluids using 50 microliters of the samples. The accuracy and reproducibility were also quite satisfactory. By use of this assay, plasma levels of NK 19 after a single oral administration of 5 mg or 10 mg/body to human volunteers or intravenous injection of 2 micrograms/kg to rats could be determined.
已开发出一种用于免疫调节剂PLATONIN（NK 19）的灵敏且特异的酶免疫测定法，并将其应用于人和大鼠的药代动力学评估。NK 19类似物琥珀酰NK 19已合成并与牛血清白蛋白偶联。该偶联物具有免疫原性，将其注射到兔体内可获得对NK 19具有高亲和力的抗体。Platonin的一些光分解产物与这些抗体的交叉反应性较低。对于竞争反应，将这些抗体与琥珀酰NK 2900-β-D-半乳糖苷酶偶联物和标准NK 19或样品的混合物一起孵育。然后，使用固定在聚苯乙烯球上的抗兔IgG将抗体结合的酶-半抗原与游离的酶-半抗原分离。通过荧光法测定固相上酶的活性。本免疫测定法使用50微升样品可检测生物流体中低至10 pg的NK 19。准确性和重现性也相当令人满意。通过使用该测定法，可以测定人志愿者单次口服5 mg或10 mg/体或大鼠静脉注射2微克/千克后NK 19的血浆水平。
