Stabilization of a chimeric malaria antigen in separation and purification through efficient inhibition of protease activity by imidazole
文献类型：期刊论文
| 作者 | Guo, Fangxia1,2; Liu, Yongdong1; Zhang, Chun1; Wang, Qi1,2; Gao, Yuhui3,4; Deng, Weiwei3,4; Wang, Heng3,4; Su, Zhiguo1,5 |
| 刊名 | PROCESS BIOCHEMISTRY
 |
| 出版日期 | 2018-02-01 |
| 卷号 | 65页码:213-219 |
| 关键词 | Multi-epitope Malaria Antigen Purification Degradation Imidazole |
| ISSN号 | 1359-5113 |
| DOI | 10.1016/j.procbio.2017.10.013 |
| 文献子类 | Article |
| 英文摘要 | 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. |
| WOS关键词 | 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 |
| 语种 | 英语 |
| WOS记录号 | WOS:000425200200026 |
| 资助机构 | 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) |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/24040]  |
| 专题 | 过程工程研究所_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 |
| 推荐引用方式 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. |
入库方式： OAI收割
来源：过程工程研究所
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