摘要
目的筛选与人重组G-CSF分子有结合活性的小分子多肽。方法将靶分子G-CSF固定在塑料平皿上,对随机噬菌体六肽库进行3轮筛选。经孵育、洗脱、扩大培养和ELISA分析,随机挑取出第3轮得到的4个阳性克隆测序,然后进行同源性比较。结果找到1个保守性肽段序列模式“SXXRVX”,此模式在人和小鼠的受体中都未见到其同源序列。结论此小肽可与G-CSF结合,故在一定程度上有可能抑制G-CSF与受体的结合,为临床治疗炎症反应和研究G-CSF与受体相互作用的机制提供了一定的帮助。
Aim To isolate small molecular polypeptides that bind to recombinant human G CSF molecule by random phage displa ying hexapeptide libraries. Methods Affinity selective method was used to screen the library. Coating rhG CSF directly on plates, hexapeptide libraries are then applied, followed by vigorous washings in detergent supplemented buffers to remove most non specifically bound phage and to select specific phage particles by the use of elutions in acid buffers. Results After three washes, four positive phage clones were selected by ELISA and sequenced. Of them, a conserved peptide motif SXXRVX was found. The amino acid sequences of these peptides were not found in the primary sequence of human and mouse G CSF′s receptors. Conclusion The peptide can bind to G CSF and may inhibit the interaction between G CSF and its receptor . This finding is helpful to provide theoretic basis for clinical treatment of inflammation and for further research on mechanism of interaction between G CSF and its receptor.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第3期287-288,共2页
Chinese Journal of Cellular and Molecular Immunology
