变形链球菌GS-5葡萄糖基转移酶B单克隆抗体及其抑制效果研究。

Pub Date : 2012-12-01 DOI:10.1089/hyb.2012.0047
Mi-Ah Kim, Min-Jeong Lee, Hae-Kyoung Jeong, Hee-Jeong Song, Hye-Jin Jeon, Kyung-Yeol Lee, Jae-Gon Kim
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引用次数: 9

摘要

变形链球菌的葡糖基转移酶-B(GTFB)被认为是一种毒力因子,因为它在产生不溶性葡聚糖方面具有活性,而不溶性葡萄糖是细菌附着在牙齿表面导致龋齿形成的关键。GTFB单克隆抗体局部被动免疫被认为是预防龋齿的有效方法。在这里我们放大了1.3 kb片段,并表达截短蛋白(GTFBN)。将表达的纯化蛋白用作BALB/c小鼠的免疫原。我们使用ELISA、斑点印迹和蛋白质印迹分析选择并建立了一种能够产生抗GTFBN的杂交瘤(HBN8)。单克隆抗GTFBN抗体通过亲和层析纯化,其同种型被确认为IgG2a。抗GTFBN抗体以剂量依赖的方式抑制变形链球菌GS-5的粗葡糖基转移酶的酶活性。这些数据表明,抗GTFBN抗体可以用作疫苗,防止变异链球菌在牙齿表面聚集,从而防止龋齿的形成。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A monoclonal antibody specific to glucosyltransferase B of Streptococcus mutans GS-5 and its glucosyltransferase inhibitory efficiency.

Glucosyltransferase-B (GTFB) of Streptococcus mutans is considered a virulence factor because of its activity in the production of insoluble glucan, which is key to the bacterial attachment onto dental surfaces, leading to the formation of dental caries. Local passive immunization with monoclonal antibodies against GTFB is considered to be an effective way to prevent dental caries. Here we amplified a 1.3 kb fragment of the N-terminal half of the gtfB gene (193-1530) of S. mutans by PCR and expressed the truncated protein (GTFBN). The expressed, purified protein was used as an immunogen in BALB/c mice. We selected and established one hybridoma (HBN8) that was capable of producing anti-GTFBN using ELISA, dot blot, and Western blot analyses. The monoclonal anti-GTFBN antibody was purified by affinity chromatography and its isotype was confirmed as IgG2a. The anti-GTFBN antibody inhibited the enzymatic activity of crude glucosyltransferase of S. mutans GS-5 in a dose-dependent manner. These data suggest that the anti-GTFBN antibody could be used as a vaccine to prevent the aggregation of S. mutans on tooth surfaces, and thus prevent the formation of dental caries.

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