Abstract
Capillary electrophoresis mass spectrometry (CE-MS) is an emerging analytical tool for microscale biological sample analysis that offers high separation resolution, low detection limit, and low sample consumption. We recently developed a novel microsampling device, “spray-capillary,” for quantitative low-volume sample extraction (as low as 15 pL/s) and online CE-MS analysis. This platform can efficiently analyze picoliter samples (e.g., single cells) with minimal sample loss and no additional offline sample-handling steps. However, our original spray-capillary-based experiments required manual manipulation of the sample inlet for sample collection and separation, which is time consuming and requires proficiency in device handling. To optimize the performance of spray-capillary CE-MS analysis, we developed an automated platform for robust, high-throughput analysis of picoliter samples using a commercially available CE autosampler. Our results demonstrated high reproducibility among 50 continuous runs using the standard peptide angiotensin II (Ang II), with an RSD of 14.70% and 0.62% with respect to intensity and elution time, respectively. We also analyzed Ang II using varying injection times to evaluate the capability of the spray-capillary to perform quantitative sampling and found high linearity for peptide intensity with respect to injection time (R2 > 0.99). These results demonstrate the capability of the spray-capillary sampling platform for high-throughput quantitative analysis of low-volume, low-complexity samples using pressure elution (e.g., direct injection). To further evaluate and optimize the automated spray-capillary platform to analyze complex biological samples, we performed online CE-MS analysis on Escherichia coli lysate digest spiked with Ang II using varying injection times. We maintained high linearity of intensity with respect to injection time for Ang II and E. coli peptides (R2 > 0.97 in all cases). Furthermore, we observed good CE separation and high reproducibility between automated runs. Overall, we demonstrated that the automated spray-capillary CE-MS platform can efficiently and reproducibly sample picoliter and nanoliter biological samples for high-throughput proteomics analysis.
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Acknowledgements
This work was partly supported by grants from OCAST HR23-169, NIH NIAID R01AI141625, and NIH NIH/NIAID2U19AI062629.
We also thank the OU Protein Production and Characterization Core (PPC) facility and Dr. Philip Bourne for supporting E. coli protein extraction. The PPC is supported by Institutional Development Awards (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health (Grants P20GM103640 and P30GM145423), the OU Vice President for Research and Partnerships, and the OU College of Arts and Sciences.
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Li, J., Huang, L., Guo, Y. et al. An automated spray-capillary platform for the microsampling and CE-MS analysis of picoliter- and nanoliter-volume samples. Anal Bioanal Chem 415, 6961–6973 (2023). https://doi.org/10.1007/s00216-023-04870-w
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DOI: https://doi.org/10.1007/s00216-023-04870-w