[en] Phage-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.
Biochemistry, biophysics & molecular biology
Author, co-author :
Fack, F.; Laboratoire National de Santé, 20A, Rue Auguste Lumière, L-1101 Luxembourg, Luxembourg
Kreis, Stephanie ; University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit
Muller, claude; CRP-Santé -Luxembourg
Heteroduplex mobility assay (HMA) pre-screening: An improved strategy for the rapid identification of inserts selected from phage-displayed peptide libraries