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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
Source PublicationPROCESS BIOCHEMISTRY
ISSN1359-5113
Volume65Pages:213-219
AbstractA 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.
KeywordMulti-epitope Malaria Antigen Purification Degradation Imidazole
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1016/j.procbio.2017.10.013
Indexed BySCI
Language英语
WOS KeywordFALCIPARUM CANDIDATE VACCINE ; VITRO PROTECTIVE EFFICACY ; IN-VITRO ; PLASMODIUM-VIVAX ; IMMUNOGENICITY ; MULTISTAGE ; STABILITY ; PARASITES ; PROTEINS ; FUSION
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering
WOS SubjectBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
Funding OrganizationNational 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 IDWOS:000425200200026
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/24040
Collection生化工程国家重点实验室
Affiliation1.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
Recommended Citation
GB/T 7714
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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