使用硝酸纤维素印迹法研究在琼脂糖凝胶中电泳的乙型肝炎表面抗原。

The use of nitrocellulose blotting for the study of hepatitis B surface antigen electrophoresed in agarose gels.

作者信息

McMichael J C, Greisiger L M, Millman I

出版信息

J Immunol Methods. 1981;45(1):79-94. doi: 10.1016/0022-1759(81)90095-8.

Abstract

Nitrocellulose-protein blotting of serum electrophoresed in agarose gels has been adapted for the study of hepatitis B surface antigen (HBsAg). 125I-labeled anti-HBs was used as the antigen probe, and the electrophoretic migration was monitored by autoradiography. The method required 3 microliter or less of serum and could detect as little as 1 pg of purified HBsAg. Typically, we observed two bands of HbsAg; a moving band which migrated about one-third the distance moved by human serum albumin and a non-migratory band which remained at the loading site. Some examples of the use of the method include: (1) empirical methods for correlating HBsAg concentration in serum to film darkness; (2) observations of mobility changes in serial sera from dialysis patients with chronic HBsAg antigenemia; and (3) detection of related antigens such as antigen from the PLC/PRF/5 hepatoma tissue culture line and the cross-reacting woodchuck patients hepatitis virus surface antigen (WHsAg).

摘要

已将琼脂糖凝胶电泳血清的硝酸纤维素蛋白印迹法应用于乙型肝炎表面抗原（HBsAg）的研究。使用125I标记的抗HBs作为抗原探针，并通过放射自显影监测电泳迁移情况。该方法所需血清量为3微升或更少，能够检测低至1皮克的纯化HBsAg。通常，我们观察到两条HBsAg条带：一条移动条带，其迁移距离约为人血清白蛋白迁移距离的三分之一；一条非迁移条带，其留在加样位点。该方法的一些应用实例包括：（1）将血清中HBsAg浓度与胶片暗度相关联的经验方法；（2）对慢性HBsAg抗原血症透析患者系列血清中迁移率变化的观察；（3）检测相关抗原，如来自PLC/PRF/5肝癌组织培养系的抗原以及交叉反应的土拨鼠肝炎病毒表面抗原（WHsAg）。