Recombinant human antibody single chain variable fragments reactive with Candida albicans surface antigens
Recombinant human antibody single chain variable fragments reactive with Candida albicans surface antigens
复制标题
DOI：
10.1016/s0022-1759(01)00463-x
复制
发表时间：
2001-11-01
影响因子：
2.2
通讯作者：
Sullivan, MA
中科院分区：
文献类型：
--
作者：
Haidaris, CG;Malone, J;Sullivan, MA
利用表达人免疫球蛋白重链和轻链可变区组合噬菌体展示文库，鉴定了能够与白色念珠菌胚分生孢子表面结合的噬菌体克隆。单链抗体可变片段（scFv）通过间接免疫荧光法（IFA）、酶联免疫吸附法（ELISA）和Western blotting检测白色念珠菌抗原。检测到的抗原在不同菌株的白色念珠菌和其他几种念珠菌中是保守的。两个scFv克隆检测到白色念珠菌胚分生孢子特异性表达的抗原；第三组在白色念珠菌的芽生孢子和丝状念珠菌中都检测到抗原。通过二硫苏糖醇可以从囊胚分生孢子中提取含有三种scFv识别的表位的抗原，表明它们通过巯基键附着在细胞壁上。scFv检测表位对高碘酸钠处理白色念珠菌芽孢孢子敏感，但对蛋白酶K不敏感，表明同源表位由碳水化合物组成。通过对皮肤念珠菌病模型的皮肤切片进行免疫组织化学染色，检测出三种scFv的抗原决定因子，证明其在体内表达。通过选择与完整生物体结合的能力，噬菌体展示系统提供了一种快速识别与念珠菌表面抗原结合的单克隆配体的方法。由于它完全是人类，因此由scFv产生的成熟抗体在治疗念珠菌病方面具有潜在的效用。(C) 2001 Elsevier Science B.V.版权所有
A combinatorial phage display library expressing human immunoglobulin heavy and light chain variable regions was used to identify phage clones capable of binding to the surface of Candida albicans blastoconidia. Single chain antibody variable fragments (scFv) derived from three clones detected C. albicans antigens by indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), and Western blotting. The antigens detected were conserved among different strains of C. albicans and several other Candida species. Two scFv clones detected antigens specifically expressed by C. albicans blastoconidia; the third detected antigens in both blastoconidia and filamentous forms of C albicans. The antigens containing the epitopes recognized by all three scFv could be extracted from blastoconidia by dithiothreitol, suggesting attachment to the cell wall via sulfhydryl bonds. Epitope detection by the scFv was sensitive to treatment of C. albicans blastoconidia with sodium periodate, but not proteinase K, indicating the cognate epitopes were composed of carbohydrate. Antigenic determinants for each of the three scFv were detected by immunohistochemical staining of skin sections from a model of cutaneous candidiasis, demonstrating expression in vivo. Through selection for the ability to bind intact organisms, the phage display system provides a means to rapidly identify monoclonal binding ligands to Candida surface antigens. Being entirely human, mature antibodies generated from the scFv have potential utility in the treatment of candidiasis. (C) 2001 Elsevier Science B.V. All rights reserved.