A new hybrid resin for stepwise screening of peptide libraries combined with single bead Edman sequencing
✍ Scribed by Hoebert S. Hiemstra; Willemien E. Benckhuijsen; Reinout Amons; Wolfgang Rapp; Jan W. Drijfhout
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 120 KB
- Volume
- 4
- Category
- Article
- ISSN
- 1075-2617
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✦ Synopsis
A library system was developed for the discovery of bioactive peptides. Library synthesis and peptide sequencing was performed on a solid support while the screening for bioactivity was done with peptides in solution. The peptides were synthesized by split and mix, one-bead-one-peptide library synthesis, using a Tentagel S-NH2 solid support with a loading of approximately 100 pmol/bead. The major part of the peptide was connected to the support by a single acid-labile linker and a minor part of the peptide was acid-stabile attached to the polymer. The percentage of acid-stabile attached peptides could easily be controlled during modification of the amino functionalities of the resin at the start of the process. The cleavage rate of the acid-labile attached peptide from the resin depends on the composition of the cleavage mixture. When cleavage conditions were carefully controlled, a three-step partial cleavage protocol allowed for convergent bioactivity screening on peptide libraries using only one type of acid-labile linker. The partial cleavage and convergent screening procedure was repeated three times, after which the bead containing the bioactive peptide was sequenced. As such a bead still contained acid-stabile attached peptide, the Edman sequencing was straightforward and repetitive yields were excellent because the immobilized peptide was not washed out.