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Easy parallel screening of reagent stability, quality control, and metrology in solid phase peptide synthesis (SPPS) and peptide couplings for microarrays

Journal Article · · Journal of Peptide Science
DOI:https://doi.org/10.1002/psc.2806· OSTI ID:1427285
 [1];  [1]
  1. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
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Evaluating the stability of coupling reagents, quality control (QC), and surface functionalization metrology are all critical to the production of high quality peptide microarrays. We describe a broadly applicable screening technique for evaluating the fidelity of solid phase peptide synthesis (SPPS), the stability of activation/coupling reagents, and a microarray surface metrology tool. This technique was used to assess the stability of the activation reagent 1-{[1-(Cyano-2-ethoxy-2-oxo-ethylidenaminooxy)dimethylamino-morpholinomethylene]}methaneaminiumHexafluorophosphate (COMU) (Sigma-Aldrich, St. Louis, MO, USA) by SPPS of Leu-Enkephalin (YGGFL) or the coupling of commercially synthesized YGGFL peptides to (3-aminopropyl)triethyoxysilane-modified glass surfaces. Coupling efficiency was quantitated by fluorescence signaling based on immunoreactivity of the YGGFL motif. It was concluded that COMU solutions should be prepared fresh and used within 5 h when stored at ~23 °C and not beyond 24 h if stored refrigerated, both in closed containers. Caveats to gauging COMU stability by absorption spectroscopy are discussed. Commercial YGGFL peptides needed independent QC, due to immunoreactivity variations for the same sequence synthesized by different vendors. This technique is useful in evaluating the stability of other activation/coupling reagents besides COMU and as a metrology tool for SPPS and peptide microarrays.
Research Organization:
Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)
Sponsoring Organization:
US Department of Homeland Security (DHS); USDOE National Nuclear Security Administration (NNSA)
Grant/Contract Number:
AC04-94AL85000
OSTI ID:
1427285
Report Number(s):
SAND--2015-6314J; 606200
Journal Information:
Journal of Peptide Science, Journal Name: Journal of Peptide Science Journal Issue: 10 Vol. 21; ISSN 1075-2617
Publisher:
Wiley - European Peptide SocietyCopyright Statement
Country of Publication:
United States
Language:
English

References (20)

Oxyma: An Efficient Additive for Peptide Synthesis to Replace the Benzotriazole-Based HOBt and HOAt with a Lower Risk of Explosion [1] journal September 2009
COMU: A Safer and More Effective Replacement for Benzotriazole-Based Uronium Coupling Reagents journal September 2009
Effect of Residual Water and Microwave Heating on the Half-Life of the Reagents and Reactive Intermediates in Peptide Synthesis journal June 2012
A Novel Family of Onium Salts Based Upon Isonitroso Meldrum's Acid Proves Useful as Peptide Coupling Reagents journal May 2010
COMU: A third generation of uronium-type coupling reagents journal January 2010
COMU: scope and limitations of the latest innovation in peptide acyl transfer reagents: COMU: SCOPE AND LIMITATIONS journal May 2013
New fluorimetric assay of horseradish peroxidase using sesamol as substrate and its application to EIA journal April 2012
An expedient conversion of α-amino acids into Weinreb amides using COMU® as a coupling agent journal January 2011
The Removal of the t -BOC Group by Electrochemically Generated Acid and Use of an Addressable Electrode Array for Peptide Synthesis journal September 2005
Morpholine-Based Immonium and Halogenoamidinium Salts as Coupling Reagents in Peptide Synthesis 1 journal April 2008
Specificity and Sensitivity of Fluorescence Labeling of Surface Species journal January 2007
On silico peptide microarrays for high-resolution mapping of antibody epitopes and diverse protein-protein interactions journal August 2012
A critical comparison of protein microarray fabrication technologies journal January 2014
Comparison of two different deposition methods of 3-aminopropyltriethoxysilane on glass slides and their application in the ThinPrep cytologic test journal January 2012
1-{[1-(Cyano-2-ethoxy-2-oxo-ethylidenaminooxy)dimethylaminomorpholinomethylene]}methane-aminium hexafluorophosphate journal July 2012
Discovery of biologically active peptides in random libraries: solution-phase testing after staged orthogonal release from resin beads. journal December 1993
Protocols for the assurance of microarray data quality and process control journal October 2005
Microarray methods and protocols: R. S. Matson, ed., CRC Press, Taylor & Francis Group, Boca Raton-London-New York, 2009, 229 p., $129.95 journal September 2010
Microarray Methods and Protocols book January 2009
TOMBU and COMBU as Novel Uronium-Type Peptide Coupling Reagents Derived from Oxyma-B journal November 2014

Cited By (1)

Synthesis of the Novel Covalent Cysteine Proteases Inhibitor with Iodoacetic Functional Group journal February 2020

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