一种用于人甲胎蛋白纯化及放射免疫测定的快速方法。

A rapid method for the purification and radioimmunoassay of human alpha-fetoprotein.

作者信息

Forrester P I, Hancock R L, Hay D M, Lai P C, Lorscheider F L

出版信息

Clin Chim Acta. 1975 Nov 3;64(3):317-23. doi: 10.1016/0009-8981(75)90360-5.

Abstract

Human alpha-fetoprotein (AFP) was isolated from cord serum on an immunoadsorbent column obtained by covalently linking rabbit anti AFP to cyanogen bromide activated Sepharose. Bound AFP was eluted with 8 M urea with better than 50% recovery. The purified AFP was iodinated prior to its use in a double antibody radioimmunoassay. The purification and radioimmunoassay employ commercially available materials. A standard inhibition curve was obtained which allowed determination of AFP levels between 50 and 100 ng/ml in human serum. The assay was verified by measureing AFP levels in normal female serum, pregnancy serum, cord serum, hepatoma ascitic fluid and a standardized AFP solution.

摘要

人甲胎蛋白（AFP）是从脐带血清中通过共价连接兔抗AFP到溴化氰活化的琼脂糖凝胶上获得的免疫吸附柱分离得到的。结合的AFP用8M尿素洗脱，回收率优于50%。纯化后的AFP在用于双抗体放射免疫测定之前进行碘化。纯化和放射免疫测定采用市售材料。获得了标准抑制曲线，可测定人血清中50至100 ng/ml的AFP水平。通过测量正常女性血清、妊娠血清、脐带血清、肝癌腹水和标准化AFP溶液中的AFP水平对该测定进行了验证。