B‐cell epitopes on the hepatitis C virus nucleocapsid protein determined by human monospecific antibodies
B‐cell epitopes on the hepatitis C virus nucleocapsid protein determined by human monospecific antibodies
复制标题
人单特异性抗体测定丙型肝炎病毒核衣壳蛋白上的 B 细胞表位
作者：
T. Akatsuka;M. Donets;L. Scaglione;W. Ching;J. Wai‐Kuo Shih;A. D. Di Bisceglie;S. Feinstone
用Epstein-巴尔病毒转化3例慢性丙型肝炎患者的B细胞，获得4株重组杆状病毒表达的抗丙型肝炎病毒核衣壳蛋白的单特异性抗体。其中一种是IgG，另外三种是IgM。它们的特异性最初通过针对具有不同丙型肝炎病毒序列插入的六种重组杆状病毒表达的丙型肝炎病毒蛋白的酶联免疫吸附试验和免疫印迹来表征。这些特异性得到了证实，并且用固相针技术制备的一系列重叠十肽更精确地确定了它们的表位。两种抗体（1F 4和2G 6）与位于核衣壳蛋白的氨基（N）末端附近的相同肽（氨基酸33-50）反应。第三种抗体（3B 5）识别由氨基酸133-142组成的肽，第四种抗体（3B 9）定位于羧基（C）末端，并与由氨基酸165-174组成的肽反应。该表位以前没有报道过。分别对核衣壳蛋白的N-末端和C-末端具有特异性的两种抗体1F 4和3B 9已稳定生产超过6个月，并正在进行亚克隆以建立单克隆性。这些抗体将成为研究丙型肝炎病毒的有用试剂。（《肝脏学》1993年;18：503-510。）
Four monospecific antibodies against the hepatitis C virus nucleocapsid protein, which was expressed by recombinant baculovirus, were obtained by Epstein‐Barr virus transformation of B cells from three patients with chronic hepatitis C virus infection. One of these antibodies was IgG and the other three were IgM. Their specificities were characterized initially by enzyme‐linked immunosorbent assay and immunoblotting against hepatitis C virus proteins expressed by six recombinant baculoviruses with different hepatitis C virus sequence insertions. These specificities were confirmed, and their epitopes were more precisely determined with a series of overlapping decapeptides made by solid‐phase pin technology. Two antibodies (1F4 and 2G6) reacted with the same peptides located near the amino(N)‐terminus of nucleocapsid protein (amino acids 33–50). The third antibody (3B5) recognized the peptide consisting of amino acids 133–142, and the fourth antibody (3B9) was mapped to the carboxy(C)‐terminus and reacted with a peptide consisting of amino acids 165–174. This epitope has not previously been reported. Two antibodies, 1F4 and 3B9, which are specific to the N‐terminus and C‐terminus of nucleocapsid protein, respectively, have been stably produced for more than 6 mo and are being subcloned to establish monoclonality. These antibodies should be useful reagents for the study of hepatitis C virus. (HEPATOLOGY 1993;18:503–510.)