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甲状旁腺素的吸附:使用放射性标记抗体进行固相测定时的陷阱?

Adsorption of parathyrin: pitfall for solid phase assays using radiolabelled antibodies?

作者信息

Jüppner H, Mohr H, Hesch R D

出版信息

J Clin Chem Clin Biochem. 1980 Sep;18(9):585-90. doi: 10.1515/cclm.1980.18.9.585.

Abstract

The physicochemical behaviour of parathyrin on surfaces was investigated. Parathyrin is bound to the wall of polyethylene, polypropylene and flint glass tubes. This adsorptive binding of the hormone, detected by labelled antibodies, is a time-dependent process which is complete within one hour. The addition of plasma decreases but does not prevent adsorption. Coating of the tubes with an anti-N-regional or anti-C-regional antiserum decreases the sensitivity of the detection of 1-34, 53-84 and 1-84 intact parathyrin, when performed in buffer. In the presence of plasma the increase of radiolabelled antibody detecting the parathyrin was more pronounced when tubes were precoated with an anti-parathyrin antibody. The sensitivity of the 1-84 parathyrin assay was also reduced when incubation was carried out in the presence of high concentrations of parathyrin-fragments, calcitonin, somatostatin or bovine serum albumin. Assay specificity seems to be related to the specificity of the labelled antibody for the respective hormonal fragment, not to that of the antibody used for coating the tubes. This investigation illustrates the necessity for the careful control of parathyrin assay conditions, e. g. non-specific binding of native parathyrin and radiolabelled tracers to laboratory reaction vessels.

摘要

研究了甲状旁腺素在表面的物理化学行为。甲状旁腺素可与聚乙烯、聚丙烯和燧石玻璃管的管壁结合。通过标记抗体检测到的这种激素的吸附性结合是一个时间依赖性过程,在一小时内完成。加入血浆会减少但不会阻止吸附。用抗N端区域或抗C端区域抗血清包被试管,在缓冲液中进行检测时,会降低检测完整的1-34、53-84和1-84甲状旁腺素的灵敏度。当试管预先用抗甲状旁腺素抗体包被时,在血浆存在的情况下,检测甲状旁腺素的放射性标记抗体的增加更为明显。当在高浓度的甲状旁腺素片段、降钙素、生长抑素或牛血清白蛋白存在下进行孵育时,1-84甲状旁腺素测定的灵敏度也会降低。测定特异性似乎与标记抗体对相应激素片段的特异性有关,而与用于包被试管的抗体的特异性无关。这项研究说明了仔细控制甲状旁腺素测定条件的必要性,例如天然甲状旁腺素和放射性标记示踪剂与实验室反应容器的非特异性结合。

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