以β-D-半乳糖苷酶为标记物的多胺抗体的制备及其在精胺和亚精胺酶免疫测定中的应用。
Preparation of polyamine antibody and its use in enzyme immunoassay of spermine and spermidine with beta-D-galactosidase as a label.
作者信息
Fujiwara K, Asada H, Kitagawa T, Yamamoto K, Ito T, Tsuchiya R, Sohda M, Nakamura N, Hara K, Tomonaga Y, Ichimaru M, Takahashi S
出版信息
J Immunol Methods. 1983 Jul 15;61(2):217-26. doi: 10.1016/0022-1759(83)90165-5.
An enzyme immunoassay for polyamines is described which uses beta-galactosidase labeled spermine and antiserum raised in rabbits against spermine-bovine serum albumin synthesized by coupling spermine to mercaptosuccinylated bovine serum albumin with a bifunctional cross-linker, N-(gamma-maleimidobutyryloxy)-succinimide. The lower limit of detection by this assay, which involves a double antibody technique for the separation of antibody-bound and free antigen, was 1 ng of spermine per tube. The anti-spermine serum showed 88% cross-reaction with spermidine but only 0.13% with putrescine, 0.08% with 1,3-diaminopropane, and 0.04% with cadaverine. The method has been used to measure serum polyamine levels in healthy subjects and cancer patients, in whom mean concentrations of 58.1 ng/ml and 94.8 ng/ml (as spermine), were respectively noted. This enzyme immunoassay is specific, accurate and easy to perform, and appears suitable for routine clinical use.
本文描述了一种用于多胺的酶免疫测定法，该方法使用β-半乳糖苷酶标记的精胺以及在兔子体内产生的抗血清，该抗血清针对通过双功能交联剂N-(γ-马来酰亚胺丁酰氧基)-琥珀酰亚胺将精胺与巯基琥珀酰化牛血清白蛋白偶联而合成的精胺-牛血清白蛋白。该测定法采用双抗体技术分离抗体结合抗原和游离抗原，其检测下限为每管1 ng精胺。抗精胺血清与亚精胺的交叉反应率为88%，但与腐胺的交叉反应率仅为0.13%，与1,3-二氨基丙烷的交叉反应率为0.08%，与尸胺的交叉反应率为0.04%。该方法已用于测量健康受试者和癌症患者的血清多胺水平，其中分别记录到平均浓度为58.1 ng/ml和94.8 ng/ml（以精胺计）。这种酶免疫测定法具有特异性、准确性且易于操作，似乎适用于常规临床应用。
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