Over-expression of fHbp in Arabdopsis for development of meningococcal serogroup B subunit vaccine.
Due to lack of commercial vaccine against the serogroup B (MenB) of Neisseria meningitides, the incidence of meningococcal disease remains high. To solve the issue, transgenic plants are used as bioreactors to produce a plant-derived fHbp subunit vaccine. In this study, the fHbp gene was optimized according to the codon usage bias of Arabidopsis thaliana, synthesized artificially, cloned into an expression vector, driven by a seed-specific promoter, and introduced into A. thaliana by Agrobacterium-mediated floral-dip transformation. Transgenic plants were identified by glufosinate selection, quickstix strips for PAT/bar tests and PCR analysis. The five plants showing higher expression of recombinant fHbp were screened through indirect ELISA. Southern blot analysis showed that the transgenic line rHF-22 had a single-copy integration and the highest expression of fHbp. Recombinant fHbp was purified from seeds of rHF-22 by nitrilotriacetic acid-mediated affinity chromatography, and the purity was 82.5%. BALB/c mice were tested for fHbp vaccine protection from lethal MenB infection, and the relative percent survival was found to be 80%. This study indicates that the recombinant fHbp produced from seeds of rHF-22 is a potential candidate for commercial MenB vaccine. It also provides a reference for safe, cheap and large-scale production of other plant-made vaccines.
College of Life Sciences, Jilin University, Changchun, China.|Wenzhou University, Wenzhou, China.
Nuo
N
Xu
Agro-Biotechnology Research Institute, Jilin Academy of Agriculture Science, Changchun, China.|Wenzhou University, Wenzhou, China.|School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, China.
Yunpeng
Y
Wang
School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, China.
Jisheng
J
Ma
Wenzhou University, Wenzhou, China.|School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, China.
Libo
L
Jin
Agro-Biotechnology Research Institute, Jilin Academy of Agriculture Science, Changchun, China.
Shaochen
S
Xing
Wenzhou University, Wenzhou, China. chaojiang10@hotmail.com.|School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, China. chaojiang10@hotmail.com.
Chao
C
Jiang
College of Life Sciences, Jilin University, Changchun, China. proflxk@163.com.|Wenzhou University, Wenzhou, China. proflxk@163.com.|School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, China. proflxk@163.com.
Xiaokun
X
Li
D000613:Aminobutyrates / Q000494:pharmacology; D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D002846:Chromatography, Affinity; D008589:Meningococcal Infections / Q000276:immunology / Q000517:prevention & control*; D022401:Meningococcal Vaccines / Q000008:administration & dosage / Q000096:biosynthesis / Q000302:isolation & purification*; D051379:Mice; D038541:Neisseria meningitidis, Serogroup B / Q000235:genetics* / Q000276:immunology; D030821:Plants, Genetically Modified / Q000254:growth & development / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000302:isolation & purification; D012639:Seeds / Q000378:metabolism; D016019:Survival Analysis
D000613:Aminobutyrates; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D022401:Meningococcal Vaccines; D011994:Recombinant Proteins; C484699:factor H-binding protein, Neisseria meningitidis; C003121:phosphinothricin
Arabidopsis thaliana; Neisseria meningitides serogroup B (MenB); Plant-derived subunit vaccine; fHbp