摘要
本试验以Cdc2相关蛋白激酶PFTK1的120～141位多肽段与载体蛋白KLH偶联制备人工抗原,免疫Balb/c小鼠,取高免疫效价的小鼠脾细胞与骨髓瘤细胞SP2/0在PEG4000作用下进行融合,通过有限稀释法最终得到了2株分泌抗PFTK1抗体的杂交瘤细胞,分别命名为4E7和2E3,经鉴定2株抗体均为IgM亚型,轻链均为κ型。经过Western blotting鉴定该抗体能够与PFTK1(1～139肽段)特异性结合。此抗体经过优球蛋白沉淀法纯化后,其纯度达到86.8%。该抗体的制备为研究蛋白激酶PFTK1功能奠定了基础。
One of PFFK1 epitopes, residues 120-141 of PFTK1, was chosen to chemically synthesize, and linked with KLH for immunization. The splenoeytes of the Balb/c with the highest serum titer was fused with myeloma cells SP2/0 by Polyethylene glycol. The two of hybridoma cells producing anti-PFTK1 monoclonal antibodies were obtained by limiting dilution, which were named 4E7 and 2E3 and whose subclass and light chain were both IgM and K type respectively. It only reacted with residues 1-139 of PFTK1 by western blot, which indicated it had good specifict for PFFK1. The ascites was purified by euglobulin precipitation and the purity of the monoclonal antibodies reached 86.8% by SDS analysis. Development of McAb may be useful for researching the function of PFTK1.
出处
《中国奶牛》
2010年第2期9-11,共3页
China Dairy Cattle
基金
国家科技支撑计划项目(2006BAD04A06)
关键词
Cdc2相关蛋白激酶
单克隆抗体
制备
Cdc2-related protein kinase PFFK1
PFFAIRE1
Monoelonal antibodies
Development
