靶向人类免疫缺陷病毒（human immunodeficiency virus，HIV）流行毒株的广谱中和抗体（broadly neutralizing antibody, bNAb）单一疗法最终会导致机体出现病毒逃逸突变，然而基于广谱中和抗体开发的双特异性或多特异性抗体则表现出较好的中和效力和广谱性。根据已公布的单链可变区基因抗体序列，经密码子优化后合成一种由单基因编码的双特异性抗体iMab-PGT151，经双酶切和测序对重组质粒进行了验证。酶联免疫吸附试验检测双特异性抗体的结合特异性，检测U87细胞裂解液中的荧光素酶活性以定量分析双特异性抗体对HIV-1假病毒的中和作用；间接免疫荧光染色法检测双特异性抗体iMab-PGT151对人喉癌上皮细胞的反应性；酶联免疫吸附试验检测该抗体对心磷脂的结合能力，验证其自体反应性。结果显示，构建的双特异抗体iMab-PGT151能够成功表达，可分别结合亲本抗体的各个配体，具有双特异性，能100%中和20株假病毒，IC50值为0.084 μg/mL。与亲本抗体相比，该抗体具有更强的中和效力和广谱度，无自体反应性，具有临床适用性。所构建的双特异性抗体iMab-PGT151将可能成为预防和治疗HIV-1感染的有效候选药物之一。

The broadly neutralizing antibody (bNAb) of human immunodeficiency virus (HIV) monotherapy eventually results in viral escape mutations in human. Bi- or multi-specific antibodies developed based on bNAbs show better neutralization potency and breadth. According to the published scFv gene antibody sequence, a bispecific antibody iMab-PGT151 encoded by a single gene was synthesized after codon optimization, and the recombinant plasmid was verified by double digestion and sequencing. Enzyme-linked immunosorbent assay (ELISA) was used to detect the binding specificity of the bispecific antibody and the luciferase activity in the lysate of U87 cells to quantitatively analyze the neutralizing effect of the bispecific antibody on the HIV-1 pseudovirus，the indirect immunofluorescence test was used to detect the reactivity of the bispecific antibody iMab-PGT151 to human laryngeal carcinoma epithelial cells and the ELISA was used to detect the antibody's ability to bind cardiolipin and verify its autoreactivity. The results showed that the constructed bispecific antibody iMab-PGT151 could be successfully expressed; it could bind each ligand of the parent antibody separately and had bispecificity; iMab-PGT151 could neutralize 100% of 20 pseudoviruses with IC50 value of 0.084 μg/mL compared with the parent antibody; the antibody had stronger neutralization potency and breath without autoreactivity, and had clinical applicability. Bispecific antibody iMab-PGT151 may become one of the promising candidates for the prevention and treatment of HIV-1 infection in the future.