Label-free protein analysis using liquid chromatography with gravimetric detection
| Autor: | Tuomas P. J. Knowles, Jerome Charmet, Tadas Kartanas, Tuuli A. Hakala, Zenon Toprakcioglu, Tom Scheidt, Aviad Levin |
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| Přispěvatelé: | Kartanas, Tadas [0000-0002-1255-4807], Levin, Aviad [0000-0002-3949-1033], Toprakcioglu, Zenon [0000-0003-1964-8432], Charmet, Jerome [0000-0001-6992-4090], Knowles, Tuomas PJ [0000-0002-7879-0140], Apollo - University of Cambridge Repository |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Detection limit
Analyte Chromatography Chemistry 010401 analytical chemistry Microfluidics Size-exclusion chromatography Fractionation Quartz crystal microbalance Biosensing Techniques 010402 general chemistry 01 natural sciences 0104 chemical sciences Analytical Chemistry Protein purification Quartz Crystal Microbalance Techniques QD Chromatography column Staphylococcal Protein A Chromatography Liquid |
| ISSN: | 0003-2700 |
| Popis: | The detection and analysis of proteins in a label-free manner under native solution conditions is an increasingly important objective in analytical bioscience platform development. Common approaches to detect native proteins in solution often require specific labels to enhance sensitivity. Dry mass sensing approaches, by contrast, using mechanical resonators, can operate in a label-free manner and offer attractive sensitivity. However, such approaches typically suffer from a lack of analyte selectivity as the interface between standard protein separation techniques and micro-resonator platforms is often constrained by qualitative mechanical sensor performance in the liquid phase. Here, we describe a strategy that overcomes this limitation by coupling liquid chromatography with a quartz crystal microbalance (QCM) platform by using a microfluidic spray dryer. We explore a strategy which allows first to separate a protein mixture in a physiological buffer solution using size exclusion chromatography, permitting specific protein fractions to be selected, desalted, and subsequently spray-dried onto the QCM for absolute mass analysis. By establishing a continuous flow interface between the chromatography column and the spray device via a flow splitter, simultaneous protein mass detection and sample fractionation is achieved, with sensitivity down to a 100 μg/mL limit of detection. This approach for quantitative label-free protein mixture analysis offers the potential for detection of protein species under physiological conditions. |
| Databáze: | OpenAIRE |
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