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  • Flow cytometric detection of proteolysis in peptide libraries synthesised on optically encoded supports.

Flow cytometric detection of proteolysis in peptide libraries synthesised on optically encoded supports.

Molecular bioSystems (2008-06-20)
Peter P T Surawski, Bronwyn J Battersby, Gwendolyn A Lawrie, Kym Ford, Andreas Rühmann, Lionel Marcon, Darby Kozak, Matt Trau
ABSTRACT

The concept of optically encoding particles for solid phase organic synthesis has existed in the literature for several years. However, there remains a significant challenge to producing particles that are capable of withstanding harsh solvents and reagents whilst maintaining the integrity and range of the optical encoding. In this study, a new generation of fluorescently encoded support particles was used for both solid phase peptide synthesis and on-particle analysis of proteolysis in a multiplexed, flow cytometric assay. The success of the assay was demonstrated through the use of a model protease, trypsin. Our results show that the use of solid supports with high peptide yield, high swellability in water and high penetration of the enzyme into the interior of the particle is not absolutely necessary for proteolysis assays.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Atto 550 azide, BioReagent, suitable for fluorescence
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Atto 488, BioReagent, suitable for fluorescence, ≥90% (HPLC)
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Atto 550 NHS ester, BioReagent, suitable for fluorescence
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Atto 620 maleimide, suitable for fluorescence
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Atto 488 azide, BioReagent, suitable for fluorescence, ≥90% (HPLC)
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Atto 488 amine, ≥90% (HPLC), BioReagent, suitable for fluorescence
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Atto 488 maleimide, BioReagent, suitable for fluorescence, ≥90% (HPLC)
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Atto 550-Biotin, BioReagent, suitable for fluorescence, ≥90% (HPLC)
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Atto 550, for fluorescence, ≥90% (HPLC)