Chang Y S, Wu C H, Chang R J, Shiuan D
Department of Biology, National Sun Yat-Sen University, Kaohsiung, Taiwan, ROC.
J Biochem Biophys Methods. 1994 Dec;29(3-4):321-9. doi: 10.1016/0165-022x(94)90042-6.
A method based on competitive enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of biotin concentrations which takes advantage of the extraordinarily high affinity between biotin and avidin. The biotin assay consisted of two steps, (i) a competition reaction between excess streptavidin-conjugated horseradish peroxidase (streptavidin-HRP) and solutions of known biotin concentrations or sample solutions and (ii) the measurement of the residual activities of the free form streptavidin-HRP which were correlated with the initial biotin concentrations. The procedure was modified by including an extra step of antibody-antigen interaction to assay biotin concentration unambiguously in more complex media. The entire assay was completed within 6 with sensitivities of approximately 1 pg/ml for biotin in a simple aqueous medium and 5 pg/ml in complex media. The method offers significant advantages in time, sensitivity and simplicity for determinations of biotin concentrations in various solutions.
开发了一种基于竞争性酶联免疫吸附测定（ELISA）的方法来测量生物素浓度，该方法利用了生物素与抗生物素蛋白之间极高的亲和力。生物素测定包括两个步骤：（i）过量的链霉亲和素偶联辣根过氧化物酶（链霉亲和素-HRP）与已知生物素浓度的溶液或样品溶液之间的竞争反应；（ii）测量与初始生物素浓度相关的游离形式链霉亲和素-HRP的残留活性。通过增加一步抗体-抗原相互作用对该程序进行了改进，以便在更复杂的介质中明确测定生物素浓度。整个测定在6小时内完成，在简单水性介质中对生物素的灵敏度约为1 pg/ml，在复杂介质中为5 pg/ml。该方法在测定各种溶液中生物素浓度的时间、灵敏度和简便性方面具有显著优势。
