Zhou G, Liu K D, Sun H C, Chen Y H, Tang Z Y, Schröder C H
Liver Cancer Institute, Shanghai Medical University, PR China.
J Cancer Res Clin Oncol. 1997;123(11-12):609-13. doi: 10.1007/s004320050113.
Monoclonal antibodies have been widely used in tumor targeting studies with promising results. However, their clinical application has been limited by heterogeneity and macro-molecular movement of murine antibody. In this study, the variable-region (heavy- and light-chain) fragments of anti-HBx monoclonal antibody were enriched by the polymerase chain reaction. The expression vector, which included a 6x histidine sequence in the 3' terminus of the HBx single-chain antibody (sFv) was recombined with a linker sequence (KLGGGGFSGA) between the variable regions. The expression product from Escherichia coli fused with 6xHis was purified by nickel (Ni2+) nitrilotriacetate chelating resin. The results of enzyme-linked immunosorbent assay and Western blotting showed that sFv had binding affinity with HBxAg, suggesting that it could become a novel targeting carrier in clinical trials.
单克隆抗体已广泛应用于肿瘤靶向研究，并取得了令人鼓舞的成果。然而，鼠源抗体的异质性和大分子移动性限制了其临床应用。在本研究中，通过聚合酶链反应富集了抗HBx单克隆抗体的可变区（重链和轻链）片段。表达载体在HBx单链抗体（sFv）的3'末端包含一个6x组氨酸序列，并在可变区之间与接头序列（KLGGGGFSGA）重组。来自大肠杆菌的与6xHis融合的表达产物通过镍（Ni2+）次氮基三乙酸螯合树脂进行纯化。酶联免疫吸附测定和蛋白质印迹结果表明，sFv与HBxAg具有结合亲和力，表明它可能成为临床试验中的新型靶向载体。
