Usuda S, Okamoto H, Tsuda F, Tanaka T, Miyakawa Y, Mayumi M
Department of Medical Sciences, Toshiba General Hospital, Tokyo, Japan.
J Virol Methods. 1998 May;72(1):95-103.
An enzyme-linked immunosorbent assay (ELISA) was developed for the determination of hepatitis B virus (HBV) core protein (p21c) using monoclonal antibody (mAb) directed to a phosphorylated C-terminal amino acid sequence that is not present in hepatitis B e antigen (HBeAg). HBV virions in the test serum were precipitated with horse polyclonal antibody to hepatitis B surface antigen (HBsAg), dissolved with Tween 20 and NaOH, and then neutralized. HBV core protein (p21c), released from HBV cores by this procedure, was sandwiched between immobilized mAb C33 directed to amino acids (aa) 133-140 of the core protein, fixed on a solid support and labeled mAb T2212 that recognizes aa 165-175, only when they are phosphorylated. The method was applied for the detection of phosphorylated p21c in sera from symptom-free carriers and patients with chronic hepatitis. The results indicated a higher extent of phosphorylation in p21c of HBV cores from symptom-free carriers than hepatitis patients.
开发了一种酶联免疫吸附测定法（ELISA），用于使用针对乙型肝炎e抗原（HBeAg）中不存在的磷酸化C末端氨基酸序列的单克隆抗体（mAb）来测定乙型肝炎病毒（HBV）核心蛋白（p21c）。用抗乙型肝炎表面抗原（HBsAg）的马多克隆抗体沉淀测试血清中的HBV病毒颗粒，用吐温20和氢氧化钠溶解，然后中和。通过该程序从HBV核心中释放的HBV核心蛋白（p21c）夹在固定在固相支持物上的针对核心蛋白氨基酸（aa）133-140的单克隆抗体C33和仅在磷酸化时识别aa 165-175的标记单克隆抗体T2212之间。该方法用于检测无症状携带者和慢性肝炎患者血清中的磷酸化p21c。结果表明，无症状携带者的HBV核心p21c中的磷酸化程度高于肝炎患者。
