脂多糖结合蛋白与CD14结合位点模拟肽的初步筛选

冯起甲; 徐智; 吴国明; 徐顺贵; 李树钧; 陈维中; 郭芮伶

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脂多糖结合蛋白与CD14结合位点模拟肽的初步筛选

论著 | 更新时间：2025-01-15

- 脂多糖结合蛋白与CD14结合位点模拟肽的初步筛选
- Preliminary screening mimic peptides of lipopolysaccharide binding protein binding to CD14
- 陆军军医大学学报 2007年29卷第16期页码：1572-1575
- 作者机构：
  
  第三军医大学新桥医院全军呼吸内科研究所, 全军呼吸病研究重点实验室
- 作者简介：
- 基金信息：
- DOI：
  中图分类号： Q503;Q516;Q755
- 纸质出版日期：2007，
  
  网络出版日期：2007-08-30，
- 稿件说明：
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冯起甲, 徐智, 吴国明, 等. 脂多糖结合蛋白与CD14结合位点模拟肽的初步筛选[J]. 陆军军医大学学报, 2007,29(16):1572-1575.

FENG QI-JIA, XU ZHI, WU GUO-MING, et al. Preliminary screening mimic peptides of lipopolysaccharide binding protein binding to CD14. [J]. Journal of army medical university, 2007, 29(16): 1572-1575.
冯起甲, 徐智, 吴国明, 等. 脂多糖结合蛋白与CD14结合位点模拟肽的初步筛选[J]. 陆军军医大学学报, 2007,29(16):1572-1575. DOI：

FENG QI-JIA, XU ZHI, WU GUO-MING, et al. Preliminary screening mimic peptides of lipopolysaccharide binding protein binding to CD14. [J]. Journal of army medical university, 2007, 29(16): 1572-1575. DOI：

摘要

目的应用噬菌体随机12肽库筛选脂多糖结合蛋白(lipopolysaccharide binding protein

LBP)与CD14结合位点的多肽序列。方法以CD14为固相筛选分子

对噬菌体12肽库进行4轮亲合筛选

采用酶联免疫吸附(ELISA)法鉴定筛选后的噬菌体克隆与CD14的结合活性，取结合活性较高的克隆进行DNA测序

推导其多肽序列并与LBP序列比对

再进行竞争抑制实验检测筛选噬菌体与LBP竞争结合CD14的效力。结果挑取的20个噬菌体克隆中，16个结合实验鉴定阳性，取6个亲和力最高的克隆测序，得到3条不同编码的12肽序列（FHRWPTWPLPSP、MHRHPPPITLPL和AAFHRAHHLTSP），3条多肽与LBP一级结构同源性低，为模拟肽。6个噬菌体克隆有较高的竞争抑制率。结论用噬菌体12肽库成功筛选出LBP与CD14结合位点的模拟肽。

Abstract

Objective To screen the mimic peptides of lipopolysaccharide binding protein (LBP) binding to CD14 from phage display random peptide library. Methods Using CD14 as a target molecule

4 rounds of biopanning to a phage random 12-mer peptide library were carried out. Twenty clones were selected randomly and used in binding test. Positive clones with high affinity were sequenced

and these peptides were compared with LBP sequence. Competitive inhibition test was carried out. Results Out of the 20 clones

16 were identified as positive. 6 of the 16 had higher affinity

and encoded 3 different 12-mer peptides such as FHRWPTWPLPSP

MHRHPPPITLPL and AAFHRAHHLTSP which were not homologous with the sequence of LBP. The phage clones with these peptides had high competitive inhibition with LBP. Conclusion Mimic peptides of LBP binding to CD14 are obtained by phage peptide library screening.

关键词

LBPCD14噬菌体肽库模拟肽

Keywords

lipopolysaccharide binding proteinCD14phage peptide librarymimic peptide

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