High expression and immunogenicity analysis of HA globular head domain of H5 subtype avian influenza virus produced in Pichia pastoris.

Wenhui Fan; Meng Wang; Lirong Liu; He Zhang; Shuang Zhang; Hongli Ling; Wenjun Liu; Jing LI

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High expression and immunogenicity analysis of HA globular head domain of H5 subtype avian influenza virus produced in Pichia pastoris.

Author: Wenhui FAN ¹ ; Meng WANG ¹ ; Lirong LIU ¹ ; He ZHANG ¹ ; Shuang ZHANG ¹ ; Hongli LING ² ; Wenjun LIU ¹ ; Jing LI ¹
Author Information

1. Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
2. Qingdao Vland Biologicals Co. Ltd., Qingdao 266114, Shandong, China.
Publication Type:Journal Article
Keywords: H5N1 influenza viruses; Pichia pastoris; hemagglutinin; subunit vaccine
MeSH: Animals; Antibodies, Viral; Birds; Hemagglutinin Glycoproteins, Influenza Virus; Influenza A Virus, H5N1 Subtype; Influenza Vaccines; Influenza in Birds; Pichia; Vaccination
From: Chinese Journal of Biotechnology 2019;35(1):49-58
CountryChina
Language:Chinese
Abstract: To evaluate the immunogenicity of HA globular head domain of H5 subtype influenza virus (H5HA), the gene of H5HA was optimized and the recombinant pPICZaA-H5HA expressing vector was constructed and transfected into Pichia pastoris. The expression of the recombinant H5HA was confirmed by SDS-PAGE and Western blotting and the results demonstrated that the recombinant H5HA (37 kDa) was highly expressed in Pichia pastoris with concentration of 0.2 mg/mL in medium. The recombinant H5HA was concentrated and purified using Ni-NTA affinity chromatography. The immunogenicity of H5HA was evaluated by immunizing eight groups of chicken through intranasal or intramuscular injection with different doses of purified H5HA combined with different adjuvants, respectively. The results showed that the recombinant H5HA could induce high level IgG (HI titer was 1:64 and neutralizing antibody titer was 1:218) and the optimal dosage of the recombinant H5HA was 50 μg combined with oil. In addition, intramuscular injection was better than nasal immunization. This study provided a theoretical support for subunit vaccine development.