A new hybrid resin for stepwise screening of peptide libraries combined with single bead Edman sequencing.
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.
Hiemstra
H S
HS
Department of Immunohematology and Blood Bank, Leiden University Hospital, The Netherlands.
Benckhuijsen
W E
WE
Amons
R
R
Rapp
W
W
Drijfhout
J W
JW
D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects; D008863:Microspheres; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D012116:Resins, Plant; D017421:Sequence Analysis / Q000379:methods*; D013268:Stimulation, Chemical
D019151:Peptide Library; D010455:Peptides; D012116:Resins, Plant