Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays
Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays
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DOI：
10.1006/meth.2000.1031
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发表时间：
2000-09-01
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影响因子：
--
通讯作者：
Butler, JE
中科院分区：
文献类型：
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作者：
Butler, JE
现代免疫分析中有很大一部分涉及使用合成固相来固定其中一种反应物。因此，这些固相免疫分析(SPI)涉及发生在靠近溶液/固相界面的反应体积内的配体-受体相互作用。因此，这些配体-受体相互作用的免疫化学/生物化学不同于它们在溶液中的对应作用。此外，固定化过程可以显著改变反应物的生物活性；大多数吸附在聚苯乙烯或硅胶上的蛋白质都被部分或大部分变性。因此，使用替代的固定化方法是有吸引力的，但可能导致总功能反应物的数量几乎没有增加。然而，所有常用的固相并不具有相同的性质或相同的反应物固定化能力，或者经历相同水平的非特异性结合。经验主义在SPIS中扮演着重要的角色。综述了用于测定固相抗体的抗原捕获、测定固相对蛋白质吸附的亲和力和估算抗体亲和力的质量定律方程的推导。(C)2000年学术出版社。
A very large proportion of modern immunoassays involve the use of synthetic solid phases to immobilize one of the reactants. These solid-phase immunoassays (SPIs) therefore involve ligand-receptor interactions that occur within a reaction volume close to the solution/solid phase interface. As a consequence, the immunochemistry/biochemistry of these ligand-receptor interactions differs from that of their counterparts in solution. Furthermore, the immobilization process can significantly alter the biological activity of the reactant; most adsorbed proteins on polystyrene or silicone are partially or largely denatured. Therefore the use of alternative methods of immobilization is attractive but may result in little increase in the amount of total functional reactant. However, all commonly used solid phases do not have the same properties or the same capacity for reactant immobilization or experience the same level of nonspecific binding. Empiricism plays a major role in SPIs. Derivations of mass law equations for measuring the antigen capture of solid-phase antibodies, for determining the affinity of solid phase for protein adsorption, and for estimating antibody affinity are reviewed. (C) 2000 Academic Press.