Characterization of native-like HIV-1 gp140 glycoprotein expressed in insect cells
| Autor: | Shaowei Li, Jun Zhang, Qingbing Zheng, Tingting Li, Yuyun Zhang, Shuangquan Gao, Zhiqing Zhang, Zekai Li, Ying Gu, Wenhui Xue, Qiaobin Yao, Honglin Shen, Zhenyong Zhang, Fang Huang, Jiaming Qiao, Rui Rong, Ningshao Xia, Shaoyong Li, Hai Yu |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Glycan
Antigenicity Insecta Glycosylation Immunogen Cell Survival Genetic Vectors 030231 tropical medicine Gene Expression HIV Antibodies Injections Intramuscular Cell Line Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Animals Humans 030212 general & internal medicine HIV vaccine Neutralizing antibody chemistry.chemical_classification Mice Inbred BALB C General Veterinary General Immunology and Microbiology biology Immunogenicity env Gene Products Human Immunodeficiency Virus Public Health Environmental and Occupational Health virus diseases Antibodies Neutralizing Virology Infectious Diseases chemistry HIV-1 biology.protein Molecular Medicine Immunization Protein Multimerization Glycoprotein Baculoviridae |
| Zdroj: | Vaccine. 37:1418-1427 |
| ISSN: | 0264-410X |
| DOI: | 10.1016/j.vaccine.2019.01.048 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect