抗hTNF-α中和抗体识别表位的筛选和鉴定

Screening and identification of anti- hTNF-α monoclonal antibody recognized hTNF-α epitopes from 7^(TM) phage display peptide library

目的:对噬菌体随机线性7肽库进行筛选,得到能与抗hTNF -α中和抗体特异性结合的表位肽. 方法: 以抗hT NF- α中和抗体为靶,筛选噬菌体线性7肽库,双夹心ELISA,竞争抑制ELISA及MAP点杂交鉴定阳性噬菌体克隆. 结果:经3轮筛选后随机挑取50个噬菌体克隆,经ELISA检测其中15个克隆显示与抗hTNF- α抗体有较强的结合能力,DNA测序结果得到的结构相似群为:WTFKMQP, WPFKMSP, WPYK MIP和WNYKMLP, 竞争性ELISA结果显示四个序列均能与TNF竞争性结合抗hTNF -αmAb,MAP点杂交结果显示筛选得到的多肽是能与抗hTNF -α抗体特异性结合的多肽. 结论:列具有很高的同源性,并能与抗hTNF -α中和抗体特异性结合,为hTNF -α相关多肽疫苗的研究提供依据.

AIM: To screen out short peptide epitopes from 7 TM phage display peptide library, which are capable of combining with anti-hTNF-α monoclonal antibody. METHODS: The epitopes of human tumor necrosis factor were screened from 7 TM phage display peptide library by using anti-hTNF-α antibody as target protein and identified by sandwich ELISA and dot-blot. RESULTS: After 3 rounds of screening, 15 of the 50 phage clones were identified as positive clones, which bound to the anti-hTNF-α antibody. The amino acid sequences of the positive clones were WTFKMQP, WPFKMSP, WPYKMIP and WNYKMLP. CONCLUSION: The framework of the screened epitopes is similar and the epitopes can specifically bind to the anti-hTNF-α antibody, which provides the basis for the development of short peptide vaccines.