Abstract:
:The efficiency of existing combinatorial biological library methods has been moderate in terms of the success rates, the affinities of the ligands selected and the time and effort involved in trying to optimize the initial leads. Although mimicking natural evolution, existing strategies take little notice of the importance of recombination within a selected population to generate increased diversity. We present an overview of our recent progress which has resulted in the successful development of such a strategy, which we designate cosmix-plexing. We incorporate recombination as a central feature in obtaining high success rates and high affinities, even for short monomer peptides, in a very short time. The method uses type II restriction enzymes to re-assort small hypervariable DNA cassettes from an intermediate pre-selected population (e.g. from a phagemid display library), while maintaining the original open-reading frame. Since, in the naive library, each cassette contains all possible combinations of the polypeptide sequences it encodes, much longer regions can be optimized than was possible with methods which depend on a simple selection from the naive library. Short peptides can now be rapidly selected, which exhibit the same, or higher, specificity and affinity for a defined target molecule, than (say) an antibody or even the natural ligand.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Collins J,Horn N,Wadenbäck J,Szardenings Mdoi
10.1016/s1389-0352(01)00019-8subject
Has Abstractpub_date
2001-06-01 00:00:00pages
317-38issue
4eissn
0168-1656issn
1873-4863pii
S1389-0352(01)00019-8journal_volume
74pub_type
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