摘要
目的 从噬菌体十二肽库中筛选结合C1q的短肽并进行初步鉴定。方法 以C1q为钓饵蛋白筛选噬菌体十二肽库 ,利用ELISA、U937细胞配体结合抑制试验、多聚IgG(AIgG)竞争抑制试验鉴定阳性克隆 ,再进行单链DNA测序和分析。结果 经 3轮筛选后 ,随机挑选 2 5个噬菌体克隆做ELISA鉴定 ,14个克隆显示与C1q有较强的结合。经过U937细胞配体结合抑制试验和AIgG竞争抑制试验鉴定后 ,将此 14个阳性克隆测序 ,从其展示肽DNA测序结果推导氨基酸序列 ,获得 9条氨基酸序列 :HWDPFSLSAYFP、WTPVRTNPFLLH、NGHLFSLSAYFP、RTQRNSPFFLCP、SPAFHPEHMGRG、SRAFHPFYRGRA、WYEGPFTLQTWP、LTQHNSPFFLLP和TSNPFFLWYPQP。结论 获得结合C1q的一些抑制性短肽 。
Objective To screen and identify C1q-binding peptides from a phage 12-mer peptide library (Ph.D.-12TM Phage Display Peptide Library). Methods The C1q-binding phage clones were screened from Ph.D.-12 library using C1q as target protein, and then identified by ELISA, U937-ligand-binding inhibitive assay, and aggregated IgG competition inhibitive assay. The sequences of single strand DNA were determined and analyzed. Results Fourteen of 25 phage clones could bind to C1q with high activity in ELISA and were positive in the two inhibitive assays. The displayed peptides of them were sequenced and the deduced amino acid sequences of selected 9 peptides are HWDPFSLSAYFP, WTPVRTNPFLLH, NGHLFSLSAYFP, RTQRNSPFFLCP, SPAFHPEHMGRG, SRAFHPFYRGRA, WYEGPFTLQTWP, LTQHNSPFFLLP, and TSNPFFLWYPQP. Conclusion C1q-binding peptide sequences with C1q-inhibiting activity are obtained, which may become the leaders of potential anti-complement therapeutic agents.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2004年第6期420-423,427,共5页
Immunological Journal
基金
国家自然科学基金 (39970 687)
广东省自然科学基金 (0 1 0 60 0 )资助项目
