禽传染性支气管炎病毒单克隆抗体的制备及其B细胞抗原表位的鉴定

为鉴定传染性支气管炎病毒(IBV)的抗原表位,本研究将IBV ck/CH/LDL/091022株免疫6周龄～8周龄BALB/c小鼠,将小鼠脾脏细胞与骨髓瘤细胞SP2/0融合,经过有限稀释法克隆筛选,得到一株针对IBV核衣壳(N)蛋白的单克隆抗体(MAb)2F2,经鉴定其染色体数目为101条,MAb亚类鉴定其重链属于IgG1,轻链属于K链.用肽扫描方法将IBV ck/CH/LDL/091022株N基因截短为具有相互部分重叠的23段,表达带GST标签的重组蛋白,与MAb 2F2进行western blot和ELISA反应后,鉴定其表位为397INWGDSAL404.将表位序列进行Blast结果表明,本研究鉴定的表位在大部分IBV株中均为保守抗原表位,为进一步建立IBV的检测方法奠定了基础.

Epitope identification of N protein in avain infectious bronchitis virus ck/CH/LDL/091022 strain

To identify the epitope of avian infectious virus(IBV),BALB/c mice of 6 to 8 week-old were immunized with IBV ck/CH/LDL/091022 strain and the spleen cells from the immunized mice were fused with mouse myeloma cells SP2/0.A positive hybridoma cell line stably secreting monoclonal antibody(MAb) was selected by 3 cycles of limited dilutions.The MAb belonged to IgG1 subtype with к chain.Epitope on IBV was identified by pepscan technique and the linear residues 397INWGDSAL404 at the C-terminus of the N protein was identified as the epitope by western blot and ELISA with the MAb.The epitope 397INWGDSAL404 is conserved in most IBV strains of different serotypes.This MAb provided a foundation for the development of diagnostic reagent in detection of IBV infection.