Quantification of Borrelia burgdorferi membrane proteins in human serum: a new concept for detection of bacterial infection

Published

November 17, 2015

Author(s)

Sao F. Cheung, Kyle Anderson, Kenia Benitez, Mark J. Soloski, John N. Aucott, Karen W. Phinney, Illarion Turko

Abstract

The low abundance of bacterial proteins in human serum upon infection imposes a challenge for the early proteomic detection of bacterial infection. To address this challenge, we propose to take advantage of detecting membrane proteins released from bacteria because of disruption of their plasma membrane triggered by the innate immune system. These membrane proteins can be separated from the bulk of serum proteins by high-speed centrifugation causing substantial sample enrichment prior to targeted protein quantification using multiple reaction monitoring mass spectrometry. This new approach was first applied to detection of B. burgdorferi membrane proteins supplemented in the human serum. The B. burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. Our results indicated that detection of B. burgdorferi membrane proteins, which are ≈107 lower in abundance than major serum proteins, is feasible. Therefore quantitative analysis was also carried out for serum samples from three patients with Lyme disease. We were able to demonstrate the detection of ospA, the major B. burgdorferi lipoprotein at the level of 4.0 fmol of ospA/mg of serum protein. The results further highlighted the concept and suggest that proposed approach can be expanded to detect other bacterial infections in humans, particularly where existing diagnostics are unreliable. 

Citation

Analytical Chemistry

Volume

87

Issue

22

Pub Type

Journals

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DOI Link

Keywords

Multiple reaction monitoring, mass spectrometry, Lyme disease, human serum

Citation

Cheung, S. , Anderson, K. , Benitez, K. , Soloski, M. , Aucott, J. , Phinney, K. and Turko, I. (2015), Quantification of Borrelia burgdorferi membrane proteins in human serum: a new concept for detection of bacterial infection, Analytical Chemistry, [online], https://doi.org/10.1021/acs.analchem.5b02803 (Accessed April 19, 2024)