TY - JOUR
T1 - Quantification of Borrelia burgdorferi Membrane Proteins in Human Serum
T2 - A New Concept for Detection of Bacterial Infection
AU - Cheung, Crystal S.F.
AU - Anderson, Kyle W.
AU - Villatoro Benitez, Kenia Y.
AU - Soloski, Mark J.
AU - Aucott, John N.
AU - Phinney, Karen W.
AU - Turko, Illarion V.
N1 - Publisher Copyright:
© 2015 American Chemical Society.
PY - 2015/10/22
Y1 - 2015/10/22
N2 - The Borrelia burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. The low abundance of bacterial proteins in human serum during infection imposes a challenge for early proteomic detection of Lyme disease. To address this challenge, we propose to detect membrane proteins released from bacteria due to 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 human serum. 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 acute 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 confirm the concept and suggest that the proposed approach can be expanded to detect other bacterial infections in humans, particularly where existing diagnostics are unreliable.
AB - The Borrelia burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. The low abundance of bacterial proteins in human serum during infection imposes a challenge for early proteomic detection of Lyme disease. To address this challenge, we propose to detect membrane proteins released from bacteria due to 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 human serum. 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 acute 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 confirm the concept and suggest that the proposed approach can be expanded to detect other bacterial infections in humans, particularly where existing diagnostics are unreliable.
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U2 - 10.1021/acs.analchem.5b02803
DO - 10.1021/acs.analchem.5b02803
M3 - Article
C2 - 26491962
AN - SCOPUS:84947282871
SN - 0003-2700
VL - 87
SP - 11383
EP - 11388
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 22
ER -