Groopman J D, Donahue P R, Zhu J Q, Chen J S, Wogan G N
Proc Natl Acad Sci U S A. 1985 Oct;82(19):6492-6. doi: 10.1073/pnas.82.19.6492.
A high-affinity IgM monoclonal antibody specific for aflatoxins was covalently bound to Sepharose 4B and used as a preparative column to isolate aflatoxin derivatives from the urine of people and experimental animals who had been exposed to the carcinogen environmentally or under laboratory conditions. Aflatoxin levels were quantified by radioimmunoassay and high-performance liquid chromatography after elution from the affinity column. In studies on rats injected with [14C]aflatoxin B1, we identified the major aflatoxin-DNA adduct, 2,3-dihydro-2-(N7-guanyl)-3-hydroxy-aflatoxin B1 (AFB1-N7-Gua), and the oxidative metabolites M1 and P1 as the major aflatoxin species present in the urine. When this methodology was applied to human urine samples obtained from people from the Guangxi Province of China exposed to aflatoxin B1 through dietary contamination, the aflatoxin metabolites detected were also AFB1-N7-Gua and aflatoxins M1 and P1. Therefore, affinity chromatography using a monoclonal antibody represents a useful and rapid technique with which to isolate this carcinogen and its metabolites in biochemical epidemiology and for subsequent quantitative measurements, providing exposure information that can be used for risk assessment.
一种对黄曲霉毒素具有高亲和力的IgM单克隆抗体与琼脂糖4B共价结合，并用作制备柱，以从环境中或实验室条件下接触过致癌物的人和实验动物的尿液中分离黄曲霉毒素衍生物。从亲和柱洗脱后，通过放射免疫分析和高效液相色谱法定量黄曲霉毒素水平。在对注射了[14C]黄曲霉毒素B1的大鼠的研究中，我们鉴定出主要的黄曲霉毒素-DNA加合物2,3-二氢-2-(N7-鸟嘌呤基)-3-羟基黄曲霉毒素B1(AFB1-N7-Gua)，以及氧化代谢产物M1和P1是尿液中存在的主要黄曲霉毒素种类。当将该方法应用于从中国广西省因饮食污染而接触黄曲霉毒素B1的人群中获得的人类尿液样本时，检测到的黄曲霉毒素代谢产物也是AFB1-N7-Gua以及黄曲霉毒素M1和P1。因此，使用单克隆抗体的亲和色谱法是一种有用且快速的技术，可用于生化流行病学中分离这种致癌物及其代谢产物，并用于后续的定量测量，提供可用于风险评估的暴露信息。
