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.

Many 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.

Adeno-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

Characterization 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.

Microdroplet 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.

A novel strategy using nanoparticle protein coronas, which enables the detection of low abundance proteins and improved plasma proteome coverage, was recently reported. Nanoparticles enable robust and reproducible measurement of the plasma proteome, which assists in the discovery of novel protein biomarkers.

Glycoproteins have been used as an essential therapeutic modality in vaccine development, with many approved clinical applications. Glycosylation of these proteins, including glycan structures and glycan occupancy, plays a vital role in stability and efficacy. EAD, a newly developed fragmentation technique unique to the ZenoTOF 7600 system (SCIEX), allows optimized peptide fragmentation in a premade platform method. This allows for accurate localization of the linked glycans alongside confident identification of the peptide through high MS/MS sequence coverage of the peptide backbone.

Product Quality (PQ) assessment of protein therapeutics by mass spectrometry is a key tool in the biopharmaceutical industry. In this study, we reported a high-throughput (HTP) mass spectrometry analysis method of non IgG proteins therapeutic direct from crude bacterial cell lysate. It allows rapid strain selection screening for confirmation of protein production.

Here we demonstrate the use of our proprietary technology, Plasma Induced Modification of Biomolecules (PLIMB), to map the epitope and paratope interactions of Trastuzumab (Herceptin™) and it’s target antigen.

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.

We 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.