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目的 研究第二抗体交连的双特异单抗聚合体对乙肝病人血液HBsAg及HBV的清除能力 ,并初步探讨其机制。方法 用羊抗鼠IgG单抗将鼠抗人CR1单抗及鼠抗HBsAg单抗连接构建成双特异单抗聚合体 (heteropolymer,HP) ,将HP加入到乙肝病人枸橼酸钠抗凝全血中 ,离心测定血浆中HBsAg及HBV的含量变化 ;设立对照组 ,并采用放射免疫、荧光定量PCR以及细胞酶联等测定方法研究细胞结合的HBsAg及HBV的含量变化及红细胞膜CR1分子数量的变化。结果 由二抗构建的HP可在 5min内将全血中 95 %以上的HBsAg及 5 1%的HBV结合到红细胞上 ,使血浆中HBsAg的含量从 30 10ng ml降到 138ng ml;HBVDNA病毒也从 7.2× 10 1 1 copies L下降到 3.5× 10 1 1 copies L。红细胞在结合抗原物质的同时伴有CR1分子的丢失 ,但红细胞无溶血反应。结论 由二抗构建的HP与文献报道的生物素 亲合素构建的HP具有相同的生物学功能 ,并更有助于对抗原物质的结合与清除 ,以红细胞为载体的HP清除系统可能具有重要的医药或临床治疗研究价值。
OBJECTIVE: To study the scavenging ability of HBsAg and HBV in blood of patients with hepatitis B due to the second antibody-linked bispecific polymer. Methods The mouse anti-human CR1 monoclonal antibody and mouse anti-HBsAg monoclonal antibody were linked by goat anti-mouse IgG monoclonal antibody to form heteropolymer (HP). HP was added to whole blood of patients with hepatitis B , The content of HBsAg and HBV in plasma were determined by centrifugation. The control group was established. The content of HBsAg and HBV and the number of CR1 in erythrocyte membrane were determined by radioimmunoassay, fluorescence quantitative PCR and enzyme-linked immunosorbent assay . Results The secondary antibody-based HP could bind more than 95% of HBsAg and 51% of HBV in whole blood to erythrocytes in 5 minutes, reducing the content of HBsAg in plasma from 30 10 ng ml to 138 ng ml. HBVDNA virus also increased from 7.2 × 10 1 1 copies L decreased to 3.5 × 10 1 1 copies L. Erythrocytes bind to antigenic substances accompanied by the loss of CR1 molecules, but no hemolytic reaction of erythrocytes. Conclusion The HP constructed by the secondary antibody has the same biological function as the HP reported in the literature of biotin-avidin, and is more helpful for the binding and clearance of antigens. The erythrocyte-based HP scavenging system may be important Medical or clinical treatment research value.