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[应用SDS-PAGE和免疫印迹法对针对问号状钩端螺旋体赖型外膜的单克隆抗体E4B7D5进行抗原分析]

[Antigen analysis of McAb E4B7D5 directed against outer envelope of Leptospira interrogans serovar lai by SDS-PAGE and immunoblot].

作者信息

Li Y, Dai B, Zhao H

出版信息

Hua Xi Yi Ke Da Xue Xue Bao. 1993 Jun;24(2):122-6.

PMID:8244286
Abstract

McAb E4B7D5 was prepared by hybridoma technology in BALB/c mice immunized to outer envelope of Leptospira interrogans serovar lai. This McAb agglutinated specifically with all the 13 serovars of Icterohaemorrhagiae serogroup in MAT test at high titres and protected the guinea pigs against the attack of virulent strain (017) of serovar lai. SDS-PAGE and immunoblot were used to analyse the reaction of the outer envelopes of the five strains of Leptospira (Leptospira interrogans Icterohaemorrhagiae serogroup, serovar lai, 017 and 601 stains; L. interrogans Hebdomadis serogroup, hebdomadis serovar strain 156; L. interrogans Australis serogroup australis serovar strain 620; biflexa leptospira patoc serovar strain Patoc I; with McAb E4B7D5. Results indicated that this McAb E4B7D5 recognized specifically 34.5 kd and 39.5 kd outer envelope proteins of serovar lai, strain 017 and 601, so it might be an immunoprotective McAb directed against Leptospira interrogans serovar lai. It was suggested that 34.5 kd and 39.5 kd outer envelope proteins of serovar lai recognized specifically by McAb E4B7D5 might be immunoprotective antigens.

摘要

用问号钩端螺旋体赖型血清群外膜免疫BALB/c小鼠,通过杂交瘤技术制备了单克隆抗体E4B7D5。该单克隆抗体在微量凝集试验中能与出血性黄疸血清群的所有13个血清型发生高滴度特异性凝集反应,并能保护豚鼠免受赖型血清型强毒株(017)的攻击。采用SDS - PAGE和免疫印迹法分析了问号钩端螺旋体的5个菌株(问号钩端螺旋体出血性黄疸血清群,赖型血清型,017和601菌株;问号钩端螺旋体七日热血清群,七日热血清型菌株156;问号钩端螺旋体澳洲血清群,澳洲血清型菌株620;双曲钩端螺旋体帕托克血清型菌株帕托克I)的外膜与单克隆抗体E4B7D5的反应。结果表明,该单克隆抗体E4B7D5能特异性识别赖型血清型、017和601菌株的34.5 kd和39.5 kd外膜蛋白,因此它可能是一种针对问号钩端螺旋体赖型血清型的免疫保护性单克隆抗体。提示单克隆抗体E4B7D5特异性识别的赖型血清型34.5 kd和39.5 kd外膜蛋白可能是免疫保护性抗原。

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