Preparation of Biocompatible Resins for Library Syntheses
By: Morten Meldal2
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The open structure of the biocompatible polyethylene glycol polyamide copolymer (PEGA resin) is presented in
Fig. 1
, exemplified by the application of the commercially available bis-2-aminopropyl-PEG1900. With the use of this PEG1900, permeability with proteins up to 50 kDa has been achieved, as demonstrated by gel permeation chromatography (1). A resin-bound fluoroescencequenched peptide substrate showed 80% cleavage with subtihsin Carlsberg (MW 27 kDa) in 1 h and
the cleavage went to completion in ∼2 h. The resin could also be used for glycopeptide assembly using bovine β-(1→4)-galactosyl
transferase (MW 43/49 kDa) to transfer galactose to the 4-position of GlcNAc (2) In this example, the diffusion and reorientation of the enzyme inside the polymer network was a rate-limiting factor for
the reaction, which could, however, be brought to completion in 72 h This indicates that the reaction was performed at the
practical limit of protein size for preparative enzyme reactions in this resin. PEGA resins perform excellently in solid-phase
assays of biomolecular reactions. Furthermore, they are transparent and no light is absorbed above 250 nm, so they can be
used with a variety of different chromophores and fluorescent probes for detection of biomolecular reactions.
Fig. 1.
Preparation of PEGA resin containing PEG1900 by the partial acryloylation procedure.
Book Title: Combinatorial Peptide Library Protocols
Series: Methods in Molecular Biology | Volume: 87 | Pub. Date: Dec-23-1997 | Page Range: 59-63 | DOI: 10.1385/0-89603-392-9:59
Subject: Protein Science
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