Agilent

Webinar Date/Time: Tuesday, January 9, 2024 at 11am EST | 8am PST | 4pm GMT| 5pm CET

Reaching sustainability and efficiency goals simultaneously is possible in RMID, a regulatory-enforced part of (bio)pharmaceutical drug manufacturing.

This technical overview discusses best handling practices for IVT mRNA and analysis with the Fragment Analyzer systems, including sample handling and quantification tips.

Webinar Date/Time: Thu, Aug 17, 2023 11:00 AM EDT

End-to-End Workflow Solutions for Oligonucleotide Analysis - From research discovery to production QA/QC

This short video provides an overview of a complete, end-to-end oligonucleotide workflow solutions

In this application note, LC separation and MS1 mass identification of a variety of oligos without the use of ion‑pairing reagents is demonstrated. The LC separation allows subsequent positive mode use with little to no flushing or hardware changes. This HILIC-based method uses an Agilent InfintyLab Poroshell 120 HILIC-Z column and MS-friendly ammonium acetate-based mobile phases. The samples were analyzed on an Agilent 1290 Infinity II LC system and a 6545XT AdvanceBio quadrupole time-of-flight mass spectrometer (LC/Q-TOF).

Characterization of oligonucleotides requires robust analytical instrumentation and methods as well as ease-of-use data analysis tools. Biocompatibility mitigates non-specific sample binding to flow path and it ensures the integrity of biomolecules and robustness of the system. In this study, two workflows, the Target Plus Impurities (TPI) and Sequence Confirmation workflows in Agilent MassHunter BioConfirm software, were carried out to characterize two oligonucleotide samples.

In this application note, the determination of oligo sequence confirmation using HILIC LC and high-resolution MS/MS data is described. As with the previous studies, an InfinityLab Poroshell 120 HILIC-Z column was used along with an Agilent 6545XT AdvanceBio LC/Q-TOF mass spectrometer.

UV-Vis spectrophotometers have been used widely for nucleic acid quantification and quality control (QC) utilizing the fact that nucleic acids have a maximum absorbance at 260 nm (1). The concentration of nucleic acids can be easily estimated using the absorbance at 260 nm and the established absorption coefficient. Often a background correction is also performed, for example collecting a baseline using a solution containing everything but the nucleic acid or by measuring the absorbance at a wavelength that nucleic acids do not absorb. Double stranded nucleic acids are bound by hydrogen bonds between the base pairs. The temperature at which double stranded nucleic acids denature to become single stranded depends on the: – sequence and length of the nucleic acid – the pH and buffer conditions – and any mismatches in base pairs between the two strands As such, the melting temperature is very useful analytical tool and can be studied by monitoring the absorbance at 260 nm as temperature is increased or decreased. As the temperature is increased, the hydrogen bonds between the strands are broken and the double stranded nucleic acid separates into two