条纹斑竹鲨的抗乙肝表面抗原的单域抗体筛选与重组抗体活性探究

The screening of Anti-HBsAgsingle domain antibody from Chiloscyllium plagiosum and the study on the activity of recombinant antibody

对免疫HBsAg-ad亚型抗原的条纹斑竹鲨中血清、脾脏和淋巴细胞进行多组学测序结果综合分析筛选出特异性序列,构建重组表达质粒pET-Duet-his-sumo-Anti-HBsAg-S1、pET-Duet-his-sumo-Anti-HBsAg-S2,分别转化大肠杆菌BL21(DE3),并诱导表达以及纯化重组单域抗体,通过ELISA和细胞水平检测重组单域抗体的活性。结果表明:通过生物信息学分析筛选出2条特异性序列,在16℃、0.5 mmol/L诱导剂条件下可获得2 mg/mL以上的重组蛋白;ELISA检测显示均与HBsAg具有较好的结合力,细胞水平活性分析得到Anti-HBsAg-S1具有较好的病毒抑制效果和生物学活性。该研究结果为其他抗原的特异性抗体筛选提供理论参考。

The serum, spleen and lymphocytes of Chiloscyllium plagiosum immune to HBsAg-adsubtype antigen were subjected to multi-omics sequencing and the specific sequences were screened from the comprehensively analyzed results. Recombinant expression plasmids pET-Duet-his-sumo-Anti-HBsAg-S1 and pET-Duet-his-sumo-Anti-HBsAg-S2(S1 and S2 represent sequences) were established and transformed into Escherichia coli BL21(DE3), respectively. Through induction expression and purification of the recombinant single-domain antibody, its activity was detected by ELISA and cell level. The results showed that two specific sequences were screened by bioinformatics analysis, and the recombinant protein above 2 mg/mL could be obtained under the effect of 16 ℃ and 0.5 mmol/L inducer. ELISA showed that all had good binding with HBsAg. Cell level activity analysis showed that anti-HBsAg-S1 had good virus inhibitory effect and biological activity. The results of thisstudy hopefully provide a theoretical reference for the screening of specific antibodies of other antigens.