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Polyethylene glycol (PEG) gel arrays for differentiating oligopeptide fragments and on-chip protease assays.
- Source :
-
Biosensors & bioelectronics [Biosens Bioelectron] 2016 Mar 15; Vol. 77, pp. 1126-33. Date of Electronic Publication: 2015 Nov 03. - Publication Year :
- 2016
-
Abstract
- Polyethylene glycol (PEG) hydrogel is permeable to biomolecules, but its permeability depends on the molecular weight of monomers and the concentration of monomer solutions. In this study, we show that PEG hydrogel made from 20% to 30% of PEG700 monomer is permeable to amino acids yet impermeable to oligopeptides. Because of its unique permeability, the gel can be used to detect protease by separating amino acids from oligopeptides when proteases cleave the oligopeptides and release amino acids. Based on this principle, an UV crosslinked gel array is fabricated on a chip for simultaneous detection of protease in up to 40 samples with only 1 µl of volume required for each sample. As a proof of concept, the on-chip protease assays are used to detect trypsin in buffer and serum. The detection limits are 1.2 nM in buffer and 17.7 nM in serum, which are comparable to conventional protease assays. Moreover, because only 1 µl of liquid is required, as little as 1.2 fmol of trypsin can be detected by using the on-chip assay. The protease assay also shows good specificity for trypsin and chymotrypsin. The gel array chip could be a useful miniaturized platform for high-throughput detection of different proteases and screening of their inhibitors.<br /> (Copyright © 2015 Elsevier B.V. All rights reserved.)
- Subjects :
- Equipment Design
Equipment Failure Analysis
Oligopeptides blood
Peptide Hydrolases chemistry
Peptide Hydrolases isolation & purification
Reproducibility of Results
Sensitivity and Specificity
Gels chemistry
Oligopeptides isolation & purification
Peptide Hydrolases blood
Polyethylene Glycols chemistry
Protein Array Analysis instrumentation
Spectrometry, Fluorescence instrumentation
Subjects
Details
- Language :
- English
- ISSN :
- 1873-4235
- Volume :
- 77
- Database :
- MEDLINE
- Journal :
- Biosensors & bioelectronics
- Publication Type :
- Academic Journal
- Accession number :
- 26569443
- Full Text :
- https://doi.org/10.1016/j.bios.2015.11.003