Flow cytometric detection of proteolysis in peptide libraries synthesised on optically encoded supports.
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.
Centre for Nanotechnology and Biomaterials, Level 5 East, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD 4072, Australia.
Peter P T
PP
Surawski
Bronwyn J
BJ
Battersby
Gwendolyn A
GA
Lawrie
Kym
K
Ford
Andreas
A
Rühmann
Lionel
L
Marcon
Darby
D
Kozak
Matt
M
Trau
D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006868:Hydrolysis; D008863:Microspheres; D017646:Organosilicon Compounds / Q000737:chemistry; D010447:Peptide Hydrolases / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*
D005456:Fluorescent Dyes; D017646:Organosilicon Compounds; D019151:Peptide Library; D010455:Peptides; D010447:Peptide Hydrolases