抗鹅IgG单克隆抗体的研制及其特性研究

Preparation and Characterization of Monoclonal Antibodies Against Goose IgG

为制备抗鹅IgG二抗用于鹅病血清学诊断，以纯化的鹅血清IgG免疫BALB／c小鼠，通过杂交瘤技术获得6株能稳定分泌抗鹅IgG的单克隆抗体（MAb）细胞株，命名为Go-1A6、Go-6A4、Go-3G11、Go-5D1、Go-688以及Go-5G5。抗体效价测定显示，细胞上清的ELISA效价为1：1600—1：4000，腹水ELISA效价为1：51200～1：204800。Westernblot分析发现，Go-5D1和Go-688与鹅IgG轻链反应，而其他4株与鹅IgG重链反应。交叉反应表明，这些MAb除了与鹅以及鸭IgG反应外，与鸡、鹌鹑、羊、牛、猪以及狗IgG不反应。间接免疫荧光试验结果显示Go-3G11和Go-6A4具有良好的反应性。结果表明，研制的MAb能为水禽疫病血清学诊断提供二抗试剂，具有一定的应用价值。

To generate secondary antibodies against goose IgG, BALB/c mice was immunized with the purified goose IgG for monoelonal antibodies （MAbs）. Through hybridomas technique, six hybridomas cell lines secreting monoclonal antibodies against goose IgG were developed, named Go-1A6, Go-6A4, Go-3G11, Go-5D1, Go-6B8 and Go-5G5. The ELISA titer of these MAbs in cell supernatant was 1 ： 1 600 to 1 ： 4 000 and that in the ascites reached to 1 ： 51 200to 1：204 800. Western blot assay revealed that MAb Go-5D1 and Go-6B8 reacted with the light chain of goose IgG, whereas Go-6A4,Go-5G5,Go-3G11 and Go-IA6 reacted with the heavy chain of goose IgG. Cross-reaction assay demonstrated that all these MAbs could react with both goose and duck IgG,but not with other IgG from chicken,quail, ovine,bovine, swine and dog,respectively. Two of these MAbs, Go-3G11 and Go-6A4,also showed good reaction for detection of the specific antibodies against goose parvovirus in indirect immunofluorescence assay. All these demonstrated that the MAbs generated here could be as secondary antibodies applied in serological diagnostics of waterfoul diseases.