Heteroduplex mobility assay (HMA) pre-screening: An improved strategy for the rapid identification of inserts selected from phage-displayed peptide libraries

被引：7

作者：

Fack, F ^{[1
]}

Deroo, S ^{[1
]}

Kreis, S ^{[1
]}

Muller, CP ^{[1
]}

机构：

[1] Lab Natl Sante, L-1101 Luxembourg, Luxembourg

来源：

MOLECULAR DIVERSITY | 2000年 / 5卷 / 01期

关键词：

DNA conformation; heteroduplex mobility assay (HMA); mismatch; mutation detection; phage display;

D O I：

10.1023/A:1011318710547

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

(P)hage-displayed peptide libraries represent an efficient tool to isolate peptides that bind a given target molecule. After several selection rounds, generally a large pool of target binding phages is obtained. Conventional analysis of the selected phage population involves extensive sequencing of many clones, most of which can be identical. We have adapted the Heteroduplex Mobility Assay (HMA) for pre-screening of phage inserts that were amplified by direct colony PCR of ELISA-positive clones. This strategy allowed for the rapid and reproducible assignment of insert sequences to different 'heteroduplex migration groups'. Sequence analysis of only one representative of each HMA migration group then completes the characterisation of the binding phage population. In our model experiments, only 16% of HMA pre-screened clones required further sequence analysis.

引用

页码：7 / 12

页数：6

共 18 条

[1] DNA-based selection and screening of peptide ligands [J].