Bioanalysis

Exogenous Molecular Analysis

During my time at Labcorp, I got the opportunity to learn about regulated bioanalysis and drug development. I worked on the LC-MS Method Development and Validation team, where I developed and validated several methods for both small and large molecules, focusing on pharmacokinetic, toxicokinetic, and therapeutic drug monitoring studies. I gained extensive hands-on experience in employing different analyte extraction strategies and utilizing liquid handling robots for high-throughput analytical workflows. These methods helped to determine therapeutic efficacies and doses, toxicity limit, dose-response relationship.

Oligonucleotide Therapeutics

We developed and validated a high-throughput LC-MS-based bioanalytical method for quantifying siRNA from mammalian biofluid and tissues. We utilized solid phase extraction and ion-pairing LC-MS/MS in this workflow.

Small Molecule Drugs

Besides nucleic acid therapeutics, I worked on developing and validating several methods for small molecule bioanalysis where I employed protein precipitation, liquid-liquid extraction, supported liquid extraction, and solid-phase extraction.

Ref.:

Boja et al. The Korean Journal of Laboratory Medicine. 2011
Nandita et al. SCIEX Application Note
Albert et al. SCIEX Application Note

Endogenous Molecular Analysis

During my graduate studies, I primarily worked with endogenous molecular LC-MS analysis, especially the metabolites that are part of bacterial cell wall biosynthesis. I also developed LC-MS methods for enzyme substrate analysis.

Chiral Analysis of Amino Acids

We developed a RPLC-MS/MS method to resolve and quantify the 20 common proteinogenic amino acid D- and L- stereoisomers using Marfey's derivatization technique. We then used the method to study these amino acids in bacterial extract. This method can be used to analyze these amino acid isomers in different biological matrix with adjustments.

MRM MS chromatogram depicting 20 proteinogenic amino acid D- and L- stereoisomers.

(Ref.: Ayon et al. JASMS 2018)

DNA Oxidative Enzyme Activity Analysis

We developed several RPLC-MS/MS methods for analyzing normal and oxidized DNA nucleosides to study a DNA oxidizing enzyme (TET2).

SDS-PAGE analysis of purified TET2 dioxygenase from E. coli BL21 (DE3) cells.

(Ref.: Bhattacharya et al. JoVE 2018)

MRM MS chromatogram for the normal and oxidized nucleosides in positive mode.

(Ref.: Bhattacharya et al. JoVE 2018)

MRM MS chromatograms of eight different nucleosides in the ion-switching mode. Period 1: positive mode (1–6 min); period 2: negative mode (6–11 min); period 3: positive mode (11–23 min).

(Ref.: Dey et al. BMP 2020)

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