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.
Department of Immunohematology and Blood Bank, Leiden University Hospital, The Netherlands.
H S
HS
Hiemstra
W E
WE
Benckhuijsen
R
R
Amons
W
W
Rapp
J W
JW
Drijfhout
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