Ion pair reversed phase LC-MS analysis of oligonucleotides presents unique challenges. By leveraging HRMS, along with software using a parsimonious deconvolution algorithm, high quality results may be obtained to better characterize impurities that are related to synthetic oligonucleotides.
Oligonucleotide Analysis by Mass Spectrometry: Sequence Display and AutomationMany oligonucleotide drug makers do not yet know what MS has to offer, and rely on sequencing, chromatography, and other assays. Here we demonstrate that MS –with appropriate software! –can assay a mixture of oligonucleotides quickly and accurately, down to trace impurities below 1% of intended product.
Comprehensive Adeno-Associated Virus (AAV) Critical Quality Attribute Analysis with LC-MSAdeno-associated viruses (AAVs) are a crucial tool for development and delivery of cutting-edge gene therapies. We developed liquid chromatography mass spectrometry (LC-MS) methods for AAV characterization for host cell protein analysis (HCP) and sequence variant analysis (SVA). In addition, orthogonal methods of subunit and peptide mapping were compared directly to capture discrepancies.
Mass spectrometric characterization glycosylated proteins present many analytical challenges Ion suppression due to the complex and heterogeneous distribution of glycans makes intact mass quantification problematic Herein we examine two common scenarios encountered in antibody characterization workflows
RapidFire-MS (RF-MS) as an Alternative to LC-MS for Rapid Assessment of Glycoforms in Therapeutic AntibodiesCharacterization of glycans is a regulatory requirement to ensure product quality and consistency between manufacturing batches. Herein, we present a comparative analysis of RapidFire; RF-MS and LC MS and demonstrate the analytical figures of merit of using RF MS for product quality screening of therapeutic proteins.
Real Time Antibody Digestion by Rapid Flow Injection Mass SpectrometryMicrodroplet reactions enable structural characterization of monoclonal antibodies at unprecedented speeds. The authors advantage of ultrafast microdroplet chemistry by coupling rapid sample introduction via flow injection to a commercial source optimized for microdroplet formation under native electrospray. They demonstrate antibody subunit analysis with
IdeS IdgE digestion, and PNGase F deglycosylation by a sandwich injection of the mAb sample between the enzymes. They apply this system for high throughput analysis of antibodies.
The S-Trap™ kit for the MAM protocol has been successfully automated on the fully integrated Tecan Freedom EVO liquid handling-Resolvex™ A200 workstation. The workflow shows efficient sample cleanup ,good peptide recovery and high reproducibility, exhibits highthroughput (upto 96 samples per run), minimizes sample processing times and removes potential operator errors, enhancing the global method performances.
Posttranslational and chemical modifications that occur during the production process of a monoclonal antibodies (mAbs) give rise to a diverse population of proteoforms with potentially differing safety, efficacy, and potency profiles. Combining icIEF with electrospray ionization (ESI) in a microfluidic chip has resulted in an analytical technique with the ability to identify and measure dozens of molecular attributes on an intact mAb in a single run.
Novel On-Chip Nebulizer for Reliable ESI andicIEF-MS BasedCharge Variant Identificationof BiopharmaceuticalsWe present a versatile microfluidic chip with on-board nebulization to facilitate stable, long lasting ESI spray, enabling reliable coupling of iCIEF to mass spectroscopy for systematic analysis of biopharmaceuticals and other biologic molecules.