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重组恶性疟原虫富含谷氨酸蛋白;纯化及其在酶联免疫吸附测定中的应用

Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay.

作者信息

Dziegiel M, Borre M B, Jepsen S, Hogh B, Petersen E, Vuust J

机构信息

Statens Seruminstitut, Copenhagen, Denmark.

出版信息

Am J Trop Med Hyg. 1991 Mar;44(3):306-13. doi: 10.4269/ajtmh.1991.44.306.

Abstract

A method for purification of a recombinant Plasmodium falciparum protein produced in E. coli and its use in an enzyme-linked immunosorbent assay (ELISA) is described. The cloned gene fragment encodes GLURP,489-1271 the carboxy-terminal 783 amino acid residue portion of a 1271 amino acid residue P. falciparum glutamate rich protein (GLURP), with a molecular weight of 220 kilodalton. The protein is associated with all parasite stages in the human host. Examination of sera from 105 adult Liberians living in a malaria endemic area revealed anti-GLURP IgG antibodies in 98% of the sera. The recombinant GLURP489-1271 was expressed as a chimeric protein, fused with E. coli beta-galactosidase. However, antibodies in sera were directed only against the malaria part of the fusion protein and not against beta-galactosidase. Antigen from in vitro P. falciparum cultures of isolates from Tanzania (F32), Papua New Guinea (MAD20) and Honduras (HB3) completely absorbed specific antibodies, indicating the presence of conserved epitopes produced by all isolates of P. falciparum. Recombinant GLURP489-1271 ELISA is sensitive and rapid, and therefore well-suited for sero-epidemiological studies, and for control of the immunogenicity of a possible future P. falciparum vaccine utilizing epitopes from GLURP.

摘要

描述了一种纯化在大肠杆菌中产生的重组恶性疟原虫蛋白的方法及其在酶联免疫吸附测定(ELISA)中的用途。克隆的基因片段编码GLURP,即1271个氨基酸残基的恶性疟原虫富含谷氨酸蛋白(GLURP)的羧基末端783个氨基酸残基部分,分子量为220千道尔顿。该蛋白与人类宿主中的所有寄生虫阶段相关。对生活在疟疾流行地区的105名成年利比里亚人的血清进行检测,发现98%的血清中存在抗GLURP IgG抗体。重组GLURP489 - 1271表达为与大肠杆菌β-半乳糖苷酶融合的嵌合蛋白。然而,血清中的抗体仅针对融合蛋白的疟疾部分,而不针对β-半乳糖苷酶。来自坦桑尼亚(F32)、巴布亚新几内亚(MAD20)和洪都拉斯(HB3)分离株的体外恶性疟原虫培养物中的抗原完全吸收了特异性抗体,表明所有恶性疟原虫分离株产生的保守表位的存在。重组GLURP489 - 1271 ELISA灵敏且快速,因此非常适合血清流行病学研究,以及用于控制未来可能利用GLURP表位的恶性疟原虫疫苗的免疫原性。

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