Quantitation of Three Outer Surface Proteins From The Lyme Disease Spirochete, Borrelia Burgdorferi By Nanoparticle Capture / Multiple Reaction Monitoring-Mass Spectrometry.

摘要

1. Trypsin digest of B. burdorferi bacterial lysate yields at least 2 suitable peptides for MRM analysis of the 3 outer surface proteins Osp A, Osp B, and Osp C. MRM shows excellent linearity from 250ng/mL to 4μg/mL of lysate spiked into urine. 2. Remazol Blue nanoparticles are effective in capturing and concentrating the 3 outer surface proteins from urine. We obtained the expected 10x concentration effect with the NPs, compared to without using the NPs. 3. MRM detection limits were at least 250ng/mL of lysate. Since the outer surface proteins are only a portion of the total amount of lysate protein, the specific protein detection limits are expected to be even lower. 4. When using the nanoparticles with 50mL of urine - vs. only 0.5mL, detection limit should theoretically be a lower by a factor of 100.