噬菌体展示三肽囊素模拟肽的研究

Studies on phage-displayed bursin-mimic peptide

本研究分离纯化抗Bursin单克隆抗体(2F9_4/HU2),利用噬菌体环7肽库筛选抗Bursin单克隆抗体(2F9_4/HU2)结合肽,测定阳性克隆ssDNA序列,并与Bursin序列进行同源性分析,通过ELISA法检测其结合活性和竞争ELISA法检测其抗原性,以Busin为对照,验证阳性克隆(№4)在鸡体内的生物学效应。序列分析表明,“LNXT”短肽序列可能为结合抗Bursin单克隆抗体(2F9_4/HU2)的保守序列,但与Bursin无同源性;噬菌体(№1)在序列中含有K、H、G。ELISA和竞争ELISA检测,表明Bursin对阳性克隆1、4、5、6号和抗Bursin单克隆抗体(2F9_4/HU2)的结合有一定的抑制作用:生物学活性初步验证表明,阳性克隆(№4)与Bursin具有类似作用,能促进抗体产生,有望成为一种模拟Bursin新型的免疫增强剂。

Anti_Bursin monoclonal antibody (2F9_4/HU2) was pruified.The peptide was biopanned with anti_Bursin monoclonal antibody (2F9_4/HU2) from phage display peptide library.The ssDNA of the positive clones were sequenced,and the binding between phage display peptide and anti_Bursin monoclonal antibody (2F9_4/HU2) was evaluated by ELISA.Antigenicity was assayed with competitive ELISA.In addition,contrast to Bursin,test was conducted to detect the bioactivity of the positive clone (No.4) in chick.“LNXT”perhaps is the conservative sequence binding with anti_Bursin monoclonal antibody (2F9_4/HU2),which is not homologous to Bursin.Positive clone (No.1) contains K、H、G,which is isogenic to Bursin.ELISA and competitive ELISA described that the positive clone 1、4、5、6 can bind well with anti_Bursin monoclonal antibody (2F9_4/HU2) and it can be competitively inhibited at certain degree by Bursin.Bioactivity test indicated that the positive phage has a similar effect on stimulating the antibody,hoping to be a new immunizing agent.