【摘要】 将猪传染性胸膜肺炎放线杆菌毒素Ⅰ(APP ApxⅠ)抗血清用饱和硫酸铵分级粗提后,再经DEAE-sepha-dex-A50低压层析系统纯化得到IgG,用此IgG作为配基对噬菌体随机十二肽库进行4轮不同条件的富集筛选。通过改变筛选条件,噬菌体的回收率从1.44×10-6增加到了8.9×10-5,P/N值逐步提高,阳性噬菌体得到了富集。ELISA结果表明,随机挑取的10个噬菌体克隆中,有5个与纯化的IgG有较强的特异性结合能力。对筛选到的5个阳性克隆提取ssDNA进行测序,并对其递呈的氨基酸序列进行分析。结果有3个克隆的4个连续的氨基酸序列(RVDV)相同,并与APP ApxⅠ蛋白的原始序列具有较高的同源性。更多还原

【Abstract】 The specific serum IgG was purified through DEAE-sephadex-A50 low pressure chromatography system after rude purification.This IgG was used as target molecular to screen the phage display 12-mer random peptide library in different conditions for 4 rounds.By altering the condition of screening,the yield ratio increased from 1.44×10^-6 to 8.9×10^-5,and the P/N value increased gradually,which meant that specific enrichment had been achieved.5 phage clones were identified as positive by ELISA and they did not have cross reactions with irrespective antibodies.ssDNA of 5 positive clones were purified for DNA sequencing and the sequences of peptides displayed on the positive clones were analyzed.4 continuous amino acid（RVDV） of 3 clones were identical,and they were homologous to the original peptide of Actinobacillus pleuropneumoniae RTX toxin Ⅰ.更多还原