Characterization of native-like HIV-1 gp140 glycoprotein expressed in insect cells

• Published in: Vaccine Vol. 37; no. 11; pp. 1418 - 1427
• Main Authors: Li, Tingting, Zhang, Zhenyong, Zhang, Zhiqing, Qiao, Jiaming, Rong, Rui, Zhang, Yuyun, Yao, Qiaobin, Li, Zekai, Shen, Honglin, Huang, Fang, Xue, Wenhui, Gao, Shuangquan, Li, Shaoyong, Zheng, Qingbing, Yu, Hai, Zhang, Jun, Gu, Ying, Li, Shaowei, Xia, Ningshao
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 07.03.2019; Elsevier Limited
• Subjects: Antigenicity; Baculovirus; Design; Envelope glycoprotein; Genes; Genetic engineering; Glycoproteins; Glycosylation; HIV; HIV-1; Human immunodeficiency virus; Immunogenicity; Immunoglobulins; Insect cell; Insect cells; Insects; Medical research; Mutation; Native-like trimer; Neutralizing; Peptides; Polysaccharides; Protein expression; Proteins; Sedimentation; Size exclusion chromatography; Trimers; Vaccine; Vaccine development; Vaccines; Viral envelope proteins; Viruses; United States > US; HIV-1; Native-like trimer; Vaccine; Insect cell; Envelope glycoprotein
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2019.01.048

•The deletion of v-cath/chiA gene enhances HIV gp140 expression yield in insect cell.•gp140 mimicked the native-like trimer and adopted distinctive glycosylation pattern.•The native-like Env proteins elicited cross-clade neutralizing antibody in mice.•The alternative gp140 production way might benefit for HIV vaccine development. The trimeric HIV-1 envelope glycoprotein (Env) is critical for vaccine development aimed at achieving broadly-neutralizing antibody responses. The use of various recombinant expression systems and construct designs are associated with the resultant nature of produced proteins, especially in terms of glycosylation, antigenicity, and immunogenicity of the glycoprotein. Here, we explored an otherwise baculovirus cassette than classical one designed to express HIV-1 Env protein, including SOSIP mutation and Foldon moiety involvement. This improved design increased the ratio of the Env trimer fraction from ∼40% to ∼60% with respect to that of prototypical design, as indicated by high-performance size-exclusion chromatography and sedimentation velocity analysis. In addition, the design prolonged cell viability and enhanced the final yield (approximately 13–15 mg/L) after affinity purification. gp140 produced from insect cells mimicked the native-like trimer and mainly adopted glycosylation pattern of oligomannose glycans. The native-like Env proteins conferred cross-clade neutralizing antibody production in BALB/c mice. In summary, the expression of Env in insect cells by optimizing the baculovirus vector provides an alternative strategy for HIV-1 immunogen production and may benefit future Env-based HIV vaccine design.