一种基于配体肽的CRP荧光检测探针的研制  

Development of A Fluorescent Probe for CRP Detection Based on Ligand Peptides

作　　者：王思雅 韩雨灏 李针保 张婷婷 梁佳欣 林卉丰 张淑华[1] WANG Siya;HAN Yuhao;LI Zhenbao;ZHANG Tingting;LIANG Jiaxin;LIN Huifeng;ZHANG Shuhua(Life Science Institute,Changchun University of Science and Technology,Changchun 130022,China)

机构地区：[1]长春理工大学生命科学技术学院,长春130022

出　　处：《生物技术进展》2022年第3期405-410,共6页Current Biotechnology

基　　金：吉林省科技厅科技发展计划项目(20210101038JC);吉林省大学生创新创业训练计划项目(2020S072)。

摘　　要：C反应蛋白(C-reaction protein,CRP)是反映机体炎症的有效标志物,早期检测是判断炎症相关疾病的关键,因此,研制CRP新型检测制剂具有重要意义。利用噬菌体表面展示技术对CRP特异性亲和配体进行了筛选,采用固相肽合成技术对目标配体进行了合成,并经生物标记、高效液相色谱法(high performance liquid chromatography,HPLC)分析及质谱(mass spectrometry,MS)鉴定,成功制得检测CRP的荧光探针。经3轮筛选、ELISA检测、重组噬菌体测序及序列比对后得到1个目标配体肽:S-P-H-N-R-S-N-L-V-Q-E-L;经肽合成及生物标记获得1种CRP荧光探针:FITC-(Acp)-S-P-H-N-R-S-N-L-V-Q-E-L。研究结果为CRP的有效检测提供了一种新型制剂。C-reactive protein(CRP)is an effective marker of inflammation in body.Early detection is the key to estimate inflam⁃mation related diseases,therefore,the development of new CRP detection preparations is of great significance.The specific affin⁃ity ligand of CRP was screened by using phage surface display techniques,and target ligands synthesized by solid⁃phase peptide synthesis technology.The fluorescent probe for CRP was successfully obtained by biomarker,high performance liquid chromatog⁃raphy(HPLC)and mass spectrometry(MS).After three rounds of screening,ELISA detection,recombinant phage sequencing and sequence alignment,a target ligand peptide was obtained:S-P-H-N-R-S-N-L-V-Q-E-L;by solid phase peptide synthesis and biomarker technology,a fluorescent probe for CRP was obtained:FITC⁃(Acp)⁃S⁃P⁃H⁃N⁃R⁃S⁃N⁃L⁃V⁃Q⁃E⁃L.This study pro⁃vided a new preparation to detect CRP effectively.

关 键 词：C反应蛋白 噬菌体表面展示技术 特异性亲和配体 酶联免疫测定分析 荧光探针 

分 类 号：Q2-33[生物学—细胞生物学] Q503