A Focus on PCR Diagnostic Probes: Oligonucleotide LC Analysis and Purification

Liquid chromatography (LC) is an indispensable method for quality control, characterization, and purification of the nucleic acids. Therapeutic oligonucleotides, mRNA vaccines, and diagnostic oligonucleotide PCR probes are typically analyzed by an ion-pairing reversed-phase LC (IP RP LC) method. Each of these application areas are readily applicable to Covid-19 research.

In this presentation we’ll discuss:

• Principles of IP RP LC
• Impact of ion-pairing system choice on oligonucleotide separation selectivity
• Purification applications for labeled oligonucleotide probes used in PCR-based diagnostics
• Applications of IP RP LC for LC MS for characterization of nucleic acids

In addition, we’ll review PREMIER solutions for the acidic nature of nucleic acids that can cause undesirable adsorption on metallic surfaces in LC systems and columns.

Presenter: Martin Gilar, Ph.D. (Scientific Fellow, Separations Research, Waters Corporation)

Dr. Martin Gilar is a Scientific Fellow in Separations Research group at Waters Corporation. He has more than 30 years of experience in the separation sciences, namely chromatography, electrophoresis, and mass spectrometry. His research interest is analysis of biopolymers such as oligonucleotides, peptides, glycans and proteins. He has published over 70 peer reviewed papers.

Dr. Gilar’s received his Ph.D. in analytical chemistry from Institute of Chemical Technology in Prague (1996). He spent postdoc years in Hybridon Inc. (1996-1998) and Northeastern University in Boston (1998) developing separation methods for antisense oligonucleotides and fraction collector for DNA molecules. Since 1998 he works at Waters Corp. in Milford, Massachusetts.