摘要
将表达伪狂犬病病毒（pseudorabies virus,PRV）gC基因的重组腺病毒经肌肉免疫BALB/c小鼠,测试了重组腺病毒免疫小鼠制备抗目的蛋白单克隆抗体的效果。ELISA分析表明,BALB/c小鼠接种重组腺病毒后能产生针对gC蛋白的特异性抗体。给BALB/c小鼠用大肠杆菌表达的gC融合蛋白MBP-gC-NC加强免疫后第3 d,取小鼠脾细胞与SP2/0小鼠骨髓瘤细胞在PEG4000作用下进行杂交融合,经筛选、克隆,获得了3株稳定分泌抗PRV gC蛋白单克隆抗体的杂交瘤细胞株,分别命名为1B4、1H4和2C8。单抗特性分析结果表明,1B4、2C8属IgG2a亚类,1H4属IgG1亚类;腹水抗体的ELISA效价为1∶2×10^5～1∶2×10^6;3株单抗都具有间接免疫荧光试验（IFA）和免疫印迹试验（Western-blotting）反应特性,能够特异地识别PRV gC蛋白。证实重组腺病毒免疫可用于制备目的蛋白单克隆抗体,为制备单抗提供了一种新的方法。
BALB/c mice were immunized intramuscularly with recombinant adenovirus expressing pseudorabies virus（PRV） glycoprotein gC gene to prepare monoclonal antibodies（McAbs） against PRV. ELISA results showed that anti-gC protein antibodies were able to be induced in immunized mice. On day 3 post-booster-immunization with the bacterially expressed gC protein MBP-gC-NC, the myeloma cell line SP2/0 was fused by PEG4000 with the splenocyte of mice after booster immunization. Then three McAbs, 1B4,1H4 and 2C8,were obtained and characterized with titers of 1∶2×10^5～1∶2×10^6in ascites. The Ig subclass of 1B4 and 2C8 belonged to IgG2a as contrast to IgG1 of 1H4. Immunofluorescence assay and Western blotting demonstrated that the McAhs were able to specifically react with PRV glycoprotein gC. The results indicated that recombinant adenovirus immunization was a novel and efficient way for preparing monoclonal antibodies against target protein.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第8期684-689,共6页
Chinese Veterinary Science
关键词
重组腺病毒
伪狂犬病病毒
gC蛋白
单克隆抗体
recombinant adenovirus
pseudorabies virus （PRV）
gC protein
monoclonal antibody
