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Stabilization of a chimeric malaria antigen in separation and purification through efficient inhibition of protease activity by imidazole
Guo, Fangxia1,2; Liu, Yongdong1; Zhang, Chun1; Wang, Qi1,2; Gao, Yuhui3,4; Deng, Weiwei3,4; Wang, Heng3,4; Su, Zhiguo1,5
2018-02-01
发表期刊PROCESS BIOCHEMISTRY
ISSN1359-5113
卷号65页码:213-219
摘要A chimeric antigen M.RCAg-1 of Plasmodium falciparum expressed in Escherichia coil was previously demonstrated inhibiting the growth of malaria parasites in vitro, but its further development has been retarded by the antigen's instability during the downstream process. In this study, it was definitely demonstrated the instability was caused by the susceptibility of M.RCAg-1 to metalloprotease(s) released from the disintegrated host cells. Interestingly, imidazole showed better inhibition effects on the degradation than EDTA. Hence, a purification procedure was successfully developed to produce M.RCAg-1 with a purity of up to 95% and an overall recovery of nearly 600 mg/L culture. When performed this protocol following the Good Manufacturing Practice regulations, the endotoxin level, the host protein content and residual DNA level, all met the FDA standards. MALDI-TOF MS demonstrated a consistent molecular weight with the theoretical value and CD revealed a mainly disordered random coil secondary structure. Immunizing mice with M.RCAg-1 with Freund's adjuvant elicited high levels of specific antibodies. Moreover, M.RCAg-1 itself could be stable at 4 degrees C for up to 6 months. Our results would provide an efficient and robust protocol for the large-scale production of M.RCAg-1 which would warrant the further development of this promising malaria vaccine candidate.
关键词Multi-epitope Malaria Antigen Purification Degradation Imidazole
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1016/j.procbio.2017.10.013
收录类别SCI
语种英语
关键词[WOS]FALCIPARUM CANDIDATE VACCINE ; VITRO PROTECTIVE EFFICACY ; IN-VITRO ; PLASMODIUM-VIVAX ; IMMUNOGENICITY ; MULTISTAGE ; STABILITY ; PARASITES ; PROTEINS ; FUSION
WOS研究方向Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering
WOS类目Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
项目资助者National Science and Technology Major Project of "National Key Program on Drug Innovation"(2013ZX09102-043) ; National Natural Science Foundation of China(21576267) ; Beijing National Science Foundation(2162041) ; Major State Basic Research Development Program of China(2013CB733604)
WOS记录号WOS:000425200200026
引用统计
文献类型期刊论文
条目标识符http://ir.ipe.ac.cn/handle/122111/24040
专题生化工程国家重点实验室
作者单位1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Chinese Acad Med Sci, Inst Basic Med Sci, Dept Microbiol & Parasitol, Beijing 100005, Peoples R China
4.Peking Union Med Coll, Sch Basic Med, Beijing 100005, Peoples R China
5.Jiangsu Natl Synerget Innovat Ctr Adv Mat SICAM, Nanjing 210023, Jiangsu, Peoples R China
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Guo, Fangxia,Liu, Yongdong,Zhang, Chun,et al. Stabilization of a chimeric malaria antigen in separation and purification through efficient inhibition of protease activity by imidazole[J]. PROCESS BIOCHEMISTRY,2018,65:213-219.
APA Guo, Fangxia.,Liu, Yongdong.,Zhang, Chun.,Wang, Qi.,Gao, Yuhui.,...&Su, Zhiguo.(2018).Stabilization of a chimeric malaria antigen in separation and purification through efficient inhibition of protease activity by imidazole.PROCESS BIOCHEMISTRY,65,213-219.
MLA Guo, Fangxia,et al."Stabilization of a chimeric malaria antigen in separation and purification through efficient inhibition of protease activity by imidazole".PROCESS BIOCHEMISTRY 65(2018):213-219.
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