Lee C Y, Chow S N, Coleman P, DeTuri M P
Biotechnol Appl Biochem. 1987 Feb;9(1):58-65.
Monoclonal antibodies specific to human prolactin were generated by an improved hybridoma technique. Following immunization with hormone conjugated with hemocyanin and cell fusion, hybrid cells were first cultured in a semisolid medium containing methylcellulose and later transferred to liquid medium for subculture. Out of 1500 colonies that were initially recovered in one single cell fusion experiment, 300 were shown to exhibit affinity to human prolactin by enzyme-linked immunosorbent assay and radioimmunoassay. Finally, nine monoclonal antibodies having high affinity and specificity to human prolactin were selected for further evaluations. From the results of a cross-matching procedure, one pair of antibodies reacting with discrete antigenic determinants of human prolactin was identified. Using a pair of monoclonal antibodies, the solid phase sandwich immunoradiometric assay and enzyme immunoassay were designed. The sensitivity of these 1-h immunoassay procedures for the determination of human prolactin was 2 ng/ml.
采用改进的杂交瘤技术制备了抗人催乳素单克隆抗体。用与血蓝蛋白偶联的激素免疫动物并进行细胞融合后，杂交细胞先在含有甲基纤维素的半固体培养基中培养，随后转移至液体培养基中进行传代培养。在一次单细胞融合实验最初获得的1500个菌落中，通过酶联免疫吸附测定法和放射免疫测定法显示有300个菌落对人催乳素表现出亲和力。最终，选择了9种对人催乳素具有高亲和力和特异性的单克隆抗体进行进一步评估。从交叉匹配实验结果中，鉴定出一对与人类催乳素不同抗原决定簇发生反应的抗体。利用这一对单克隆抗体，设计了固相夹心免疫放射测定法和酶免疫测定法。这些用于测定人催乳素的1小时免疫测定法的灵敏度为2 ng/ml。
