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抗曼氏血吸虫人尿蛋白Sm-UP(2)IP的小鼠单克隆抗体。

Mouse monoclonal antibodies against human urinary protein Sm-UP(2)IP of Schistosoma mansoni.

作者信息

Boamah D, Ayi I, Yankson K, Bosompem K M

机构信息

Institute of Tropical Medicine, Nagasaki University, Sakamoto, Nagasaki City, Japan.

出版信息

Hybridoma (Larchmt). 2012 Jun;31(3):188-95. doi: 10.1089/hyb.2011.0105.

Abstract

There are innumerable clinical and pathological problems associated with schistosomiasis that have necessitated various control programs. Successful control would naturally depend on effective rapid diagnosis in the field. However, the overlapping distribution of urinary and intestinal schistosomiasis in hyperendemic areas calls for differential diagnosis. This study was aimed at producing anti-Schistosoma mansoni monoclonal antibodies (MAbs) for possible utilization in assays to detect antigens in the urine of infected persons. In order to raise antibodies to less immunogenic urinary parasite antigens, BALB/c mice were immunized with Schistosoma mansoni soluble worm antigens (Sm-SWA) while urinary proteins (Sm-UP(2)IP), isolated from infected human urine samples, was used as a final booster before cell fusion. Hybridoma cells were obtained by the fusion of mouse myeloma and spleen cells from the immunized mice, which were screened by microplate ELISA and then studied further to obtain anti-S. mansoni specific MAbs. The MAbs analyzed presented IgM isotypes. The reactivity of anti-S. mansoni MAbs with Sm-UP(2)IP, 13/43 (30.2%), MAbs showed stronger reactivity. It was observed that one of the MAbs cross-reacted with antigen associated with S. haematobium urinary antigen (Sh-UP(2)IP). Nine (9/13, 69.2%) MAbs recognized glycoprotein antigenic epitopes of Sm-UP(2)IP and Sm-SWA. On the other hand, 4/13 (30.8%) MAbs recognized carbohydrate antigenic epitopes. Band size of 8.9 kDa associated with Sm-UP(2)IP was detected by the 13 MAbs. With Sm-SWA, all the MAbs detected band sizes of 177.8 and 158.5 kDa. In addition, three MAbs recognized a 38.9 kDa band. The generation of anti-S. mansoni species-specific MAbs offers opportunities to develop a specific MAb-based diagnostic tool for use in the field to detect Schistosoma mansoni infection in Ghana.

摘要

血吸虫病存在无数与临床和病理相关的问题,这使得各种控制计划成为必要。成功的控制自然取决于现场有效的快速诊断。然而,在高度流行地区,泌尿生殖系统血吸虫病和肠道血吸虫病的分布重叠,需要进行鉴别诊断。本研究旨在制备抗曼氏血吸虫单克隆抗体(MAb),以便可能用于检测感染者尿液中抗原的检测方法。为了产生针对免疫原性较低的泌尿寄生虫抗原的抗体,用曼氏血吸虫可溶性虫体抗原(Sm-SWA)免疫BALB/c小鼠,同时将从感染人类尿液样本中分离出的尿蛋白(Sm-UP(2)IP)用作细胞融合前的最终加强免疫。通过将小鼠骨髓瘤细胞与免疫小鼠的脾细胞融合获得杂交瘤细胞,通过微孔板ELISA进行筛选,然后进一步研究以获得抗曼氏血吸虫特异性单克隆抗体。分析的单克隆抗体呈现IgM同种型。抗曼氏血吸虫单克隆抗体与Sm-UP(2)IP的反应性,13/43(30.2%)的单克隆抗体显示出更强的反应性。观察到其中一种单克隆抗体与埃及血吸虫尿抗原(Sh-UP(2)IP)相关抗原发生交叉反应。9种(9/13,69.2%)单克隆抗体识别Sm-UP(2)IP和Sm-SWA的糖蛋白抗原表位。另一方面,4/13(30.8%)单克隆抗体识别碳水化合物抗原表位。13种单克隆抗体检测到与Sm-UP(2)IP相关的8.9 kDa条带大小。对于Sm-SWA,所有单克隆抗体检测到的条带大小为177.8和158.5 kDa。此外,3种单克隆抗体识别38.9 kDa条带。抗曼氏血吸虫种特异性单克隆抗体的产生为开发一种基于单克隆抗体的特异性诊断工具提供了机会,该工具可用于现场检测加纳的曼氏血吸虫感染。

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