Nyalala J O, Livaniou E, Leondiadis L, Evangelatos G P, Ithakissios D S
Radioimmunochemistry Lab., NCSR Demokritos, Athens, Greece.
J Immunoassay. 1997 Feb;18(1):1-19. doi: 10.1080/01971529708005802.
An indirect enzyme-linked assay was developed for quantifying biotin concentrations in human sera. Biotin standard solutions or unknown samples are preincubated with streptavidin-conjugated horseradish peroxidase (streptavidin-HRP) and added to plates coated with biotinylated bovine IgG (B-IgGb). The concentration of the streptavidin-HRP is such that the streptavidin binding sites are sufficient to bind apparently all the biotin present in samples, whereas, the remaining sites are inversely proportional to the amount of biotin in analysed sample. These sites could subsequently interact with the immobilized B-IgGb providing signal. The assay demonstrated dynamic range 5 to 640 ng/L, detection limit 2 ng/L, intra- and interassay C.V., 1.6-3.9% and 3.7-7.2% respectively, recovery 100-114% and linear recovery 90-117%. Serum biotin determined: healthy individuals 66 to 600 ng/L, pregnant women (> or = 36 weeks) 60 to 360 ng/L, and patients under chronic haemodialysis 0.56 to 1.62 micrograms/L. The method described is among those few which have been experimentally evaluated for their capabilitity of assessing biotin in human sera.
开发了一种间接酶联测定法，用于定量人血清中的生物素浓度。将生物素标准溶液或未知样品与链霉亲和素偶联的辣根过氧化物酶（链霉亲和素 - HRP）预孵育，然后加入包被有生物素化牛IgG（B - IgGb）的平板中。链霉亲和素 - HRP的浓度应使链霉亲和素结合位点足以结合样品中明显存在的所有生物素，而其余位点与分析样品中生物素的量成反比。这些位点随后可与固定化的B - IgGb相互作用产生信号。该测定法的动态范围为5至640 ng/L，检测限为2 ng/L，批内和批间变异系数分别为1.6 - 3.9%和3.7 - 7.2%，回收率为100 - 114%，线性回收率为90 - 117%。测定的血清生物素水平为：健康个体66至600 ng/L，孕妇（≥36周）60至360 ng/L，慢性血液透析患者0.56至1.62 μg/L。所描述的方法是少数经过实验评估其评估人血清中生物素能力的方法之一。
