Performance comparison of three trypsin columns used in liquid chromatography

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F17%3A10368328" target="_blank" >RIV/00216208:11310/17:10368328 - isvavai.cz</a>
Result on the web
<a href="https://doi.org/10.1016/j.chroma.2017.02.024" target="_blank" >https://doi.org/10.1016/j.chroma.2017.02.024</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.chroma.2017.02.024" target="_blank" >10.1016/j.chroma.2017.02.024</a>

Result language
angličtina
Original language name
Performance comparison of three trypsin columns used in liquid chromatography
Original language description
Trypsin is the most widely used enzyme in proteomic research due to its high specificity. Although the in solution digestion is predominantly used, it has several drawbacks, such as long digestion times, autolysis, and intolerance to high temperatures or organic solvents. To overcome these shortcomings trypsin was covalently immobilized on solid support and tested for its proteolytic activity. Trypsin was immobilized on bridge-ethyl hybrid silica sorbent with 300 (A) over circle pores, packed in 2.1 x 30mm column and compared with Perfinity and Poroszyme trypsin columns. Catalytic efficiency of enzymatic reactors was tested using N alpha-Benzoyl-L-arginine 4-nitroanilide hydrochloride as a substrate. The impact of buffer pH, mobile phase flow rate, and temperature on enzymatic activity was investigated. Digestion speed generally increased with the temperature from 20 to 37 degrees C. Digestion speed also increased with pH from 7.0 to 9.0; the activity of prototype enzyme reactor was highest at pH 9.0, when it activity exceeded both commercial reactors. Preliminary data for fast protein digestion are presented.
Czech name
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Czech description
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Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10406 - Analytical chemistry

Project
<a href="/en/project/GA16-05942S" target="_blank" >GA16-05942S: Gradient chromatofocusing and isoelectric focusing - tools for separation of proteins</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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