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10.3390/ijms23042307
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Article

Comparison of Different Mass Spectrometry Workflows for the Proteomic Analysis of Tear Fluid

by
Garrett Jones
1,
Tae Jin Lee
1,
Joshua Glass
1,
Grace Rountree
1,
Lane Ulrich
2,
Amy Estes
2,
Mary Sezer
2,
Wenbo Zhi
1,
Shruti Sharma
1,2 and
Ashok Sharma
1,2,3,*
1
Center for Biotechnology and Genomic Medicine, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA
2
Department of Ophthalmology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA
3
Department of Population Health Sciences, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2022, 23(4), 2307; https://doi.org/10.3390/ijms23042307
Submission received: 23 December 2021 / Revised: 1 February 2022 / Accepted: 15 February 2022 / Published: 19 February 2022
(This article belongs to the Special Issue Emerging OMICS Approaches to Studying the Eye in the Norm and Pathology 2.0)
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Abstract

The tear film is a multi-layer fluid that covers the corneal and conjunctival epithelia of the eye and provides lubrication, nutrients, and protection from the outside environment. Tear fluid contains a high concentration of proteins and has thus been recognized as a potential source of biomarkers for ocular disorders due to its proximity to disease sites on the ocular surface and the non-invasive nature of its collection. This is particularly true in the case of dry eye disease, which directly impacts the tear film and its components. Proteomic analysis of tear fluid is challenging mainly due to the wide dynamic range of proteins and the small sample volumes. However, recent advancements in mass spectrometry have revolutionized the field of proteomics enabling unprecedented depth, speed, and accuracy, even with small sample volumes. In this study using the Orbitrap Fusion Tribrid mass spectrometer, we compared four different mass spectrometry workflows for the proteomic analysis of tear fluid collected via Schirmer strips. We were able to establish a method of in-strip protein digestion that identified >3000 proteins in human tear samples from 11 healthy subjects. Our method offers a significant improvement in the number of proteins identified compared to previously reported methods without pooling samples.
Keywords: tear film; proteomics; biomarkers; dry eye disease; ocular surface; mass spectrometry tear film; proteomics; biomarkers; dry eye disease; ocular surface; mass spectrometry

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MDPI and ACS Style

Jones, G.; Lee, T.J.; Glass, J.; Rountree, G.; Ulrich, L.; Estes, A.; Sezer, M.; Zhi, W.; Sharma, S.; Sharma, A. Comparison of Different Mass Spectrometry Workflows for the Proteomic Analysis of Tear Fluid. Int. J. Mol. Sci. 2022, 23, 2307. https://doi.org/10.3390/ijms23042307

AMA Style

Jones G, Lee TJ, Glass J, Rountree G, Ulrich L, Estes A, Sezer M, Zhi W, Sharma S, Sharma A. Comparison of Different Mass Spectrometry Workflows for the Proteomic Analysis of Tear Fluid. International Journal of Molecular Sciences. 2022; 23(4):2307. https://doi.org/10.3390/ijms23042307

Chicago/Turabian Style

Jones, Garrett, Tae Jin Lee, Joshua Glass, Grace Rountree, Lane Ulrich, Amy Estes, Mary Sezer, Wenbo Zhi, Shruti Sharma, and Ashok Sharma. 2022. "Comparison of Different Mass Spectrometry Workflows for the Proteomic Analysis of Tear Fluid" International Journal of Molecular Sciences 23, no. 4: 2307. https://doi.org/10.3390/ijms23042307

APA Style

Jones, G., Lee, T. J., Glass, J., Rountree, G., Ulrich, L., Estes, A., Sezer, M., Zhi, W., Sharma, S., & Sharma, A. (2022). Comparison of Different Mass Spectrometry Workflows for the Proteomic Analysis of Tear Fluid. International Journal of Molecular Sciences, 23(4), 2307. https://doi.org/10.3390/ijms23042307

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