On Chip Two-Dimensional Separation Device utilizing Radical Activated Cleavage (RAC) for the Fragmentation of Biological Samples

Mark Hayes (Inventor)

Research output: Patent

Abstract

Tryptic digests with the subsequent analysis of the fragments produced are frequently used to characterize proteins. Such digests are limited by the amino acid specificity of the digesting enzyme (K and R in the case of trypsin). The analysis of fragments may also be complicated by self-digestion of the enzyme. The present invention provides a method for the digestion of proteins and other biomolecules by the in situ production of hydroxyl radicals from an excited semiconductor source. The concentration of the radicals, and hence the degree of digestion, can be precisely tuned and is highly reproducible. The methodology can be integrated into microfluidic and HPLC systems for the analysis of single proteins or complex mixtures. Modifications of the system may allow for the analysis of other biomolecules such as DNA and RNA. Potential Applications Protein characterization and identification - based on distinct fragmentation pattern observed Modifications may allow for nucleic acid characterizationBenefits and Advantages Analytic Unique, reproducible digestion patterns can be obtained. Integration with microfluidics reduces sample requirements and limits chemical waste. Hydroxyl RAC User Friendly Platform There is no need for enzyme storage or titration. The integration of sample digestion with analysis reduces steps and improves reproducibility of analysis. Highly Tunable - Cleavage parameters can be modified by increasing or decreasing hydroxyl radical production at the semiconductor surface. Clearer, Easier to Interpret Results No need for an added endopeptidase or other enzyme lowers cost, simplifies procedures and translates into results that are easier to interpret. Download original PDFFor more information about the inventor(s) and their research, please see Dr. Hayes' departmental webpageDr. Hayes' research webpage
Original languageEnglish (US)
StatePublished - Mar 12 2003

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