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Infectious agents like viruses and bacteria pose a continuing menace to human health. The clear
understanding of their roles as friends or foes depends on identification of factors that guide their
infectivity. Therefore, bioanalytical assays that throw light on these factors are in increasing need.
In my work, I have developed tools using mass spectrometry and fluorescence-based techniques
to probe into bacterial and viral infectivity.
I have divided my presentation into three parts.
1) The turn-over of bacterial LpxC (UDP-3-O-acyl-N-acetylglucosamine deacetylase) in
determining sustained drug effect: use of SILAC as a tool
Target vulnerability dictates dosing requirements, with high vulnerability targets requiring less
frequent dosing. However, the turn-over of the target (the resynthesis or degradation) can
modulate the target-vulnerability and thereby the advantages of a long-residence time inhibitor as
well. The turn-over of the target has been demonstrated using pulsed SILAC (Stable Isotope
Labelling by Amino acids in Cell culture) in a mutant and wild-type strain of E. coli.
2) LC-MS Assay for Quantification of Proteins in Oncolytic vesicular stomatitis virus (VSVGP)
In the second part of the talk, LC MS/MS using multiple reaction monitoring (MRM) has been
leveraged to study a new drug modality called oncolytic virus (OV)- Vesicular Stomatitis Virus
(VSV). Our results demonstrate that this assay can provide absolute quantification of the viral
proteins.
3) The effect of small extracellular vesicles on infection by Respiratory Syncytial Virus (RSV):
Cellular tropism
In the third part of my talk, the infectious ability of Respiratory Syncytial Viruses (RSV) has been
investigated in response to small extracellular vesicles. This work investigates the tropism
between cell lines in response to infection by RSV in the presence of vesicles using
immunofluorescence, cell-based assays, q-RTPCR and Western blotting as techniques. |