[抗H9亚型禽流感病毒血凝素单克隆抗体的制备及鉴定]

[Preparation and characterization of monoclonal antibodies against the hemagglutinin of H9 subtype of avian influenza virus].

作者信息

Zeng Yan-bing, Jiao Xin-an, Pan Zhi-ming, Huang Jin-lin, Zhang Ping-hu, Zhang Su-hua, Sun Quan-yun, Liu Xiu-fan

机构信息

Key Laboratory of Animal Lemology, Ministry of Agriculture/College of Animal Science and Veterinary Medicine, Yangzhou University 225009, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Nov;20(6):702-4.

PMID:15555440

Abstract

AIM

To prepare monoclonal antibodies (mAb) against the hemagglutinin(HA) of H9 subtype of avian influenza virus (AIV).

METHODS

8 week-old female BALB/c mice were immunized with the inactivated vaccine of H9 subtype of AIV. Splenocytes from the immunized mice were fused with Sp2/0 myeloma cells, and positive hybridoma clones were screened by indirect ELISA and hemagglutination inhibition test (HI). The specificity of the mAb was characterized by ELISA, HI test, indirect immunofluorescence (IF) staining and Western blot.

RESULTS

Three hybridoma cell lines named 2H1, 2A3 and 1C8 against HA of AIV H9 were obtained. The HI titers of 3 mAbs were 1 x 2(8)-1 x 2(13), and the ELISA titers were 1 x 10(-7), 1 x 10(-5) and 5 x 10(-6), respectively. The immunoglobulin subclass of all 3 mAbs was IgG1. Western blot analysis confirmed that mAb 2H1 could recognize HA and reacted to 31 out of 32 isolates of H9 subtype of AIV.

CONCLUSION

Three mAbs recognizing HA of H9 subtype of AIV were obtained, which may provide an useful tool for the antigenic analysis, the serological diagnosis, the epidemiological survey and the evaluation of AIV vaccine.

摘要

目的

制备抗H9亚型禽流感病毒（AIV）血凝素（HA）的单克隆抗体（mAb）。

方法

用H9亚型AIV灭活疫苗免疫8周龄雌性BALB/c小鼠。将免疫小鼠的脾细胞与Sp2/0骨髓瘤细胞融合，通过间接ELISA和血凝抑制试验（HI）筛选阳性杂交瘤克隆。通过ELISA、HI试验、间接免疫荧光（IF）染色和蛋白质印迹法对mAb的特异性进行鉴定。

结果

获得了3株针对AIV H9 HA的杂交瘤细胞系，分别命名为2H1、2A3和1C8。3株mAb的HI效价为1×2⁸-1×2¹³，ELISA效价分别为1×10⁻⁷、1×10⁻⁵和5×10⁻⁶。3株mAb的免疫球蛋白亚类均为IgG1。蛋白质印迹分析证实mAb 2H1能够识别HA，并与32株H9亚型AIV中的31株发生反应。