Development and optimization of a TNFα neutralizing assay using a Quality by Design approach

Development and optimization steps are of major importance to ensure the delivery of reliable results in the routine use of a bioassay. Quality by Design (QbD) approach requires a combination of thorough knowledge of the scientific background of the assay and the use of proper statistical tools to achieve best results.

Absolute quantification of proteins & mAbs by ICP-MS

Although drug products issued from biotechnology have now been marketed for many years, accurate protein quantifcation remains a challenge.

State-of-the art LC/MS methods applied to the characterisation of a highly-glycosylated fusion protein: Etanercept

A large number of pharmaceuticals are glycosylated proteins. An adequate glycosylation is critical for therapeutic proteins in terms of safety, bioactivity, solubility, stability, and pharmacokinetics and dynamics.

Multi-level mass spectrometric characterisation of Antibody-Drug Conjugates in a regulated environment.

Antibody-Drug Conjugates (ADCs) are antibodies engineered to deliver a cytotoxic agent specifically to tumor cells expressing a defined antigen.

Development and use of 2D-LC/MS as a versatile and powerful tool for the analysis of mAbs and ADCs in a regulated environment.

In the recent years, monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) have emerged as potent and specific anticancer agents. However, due to their size, complexity, and heterogeneity, their physico-chemical characterisation requires a large number of assays aimed at verifying their sequence identity, post-translational modifications, and – in the case of ADCs – range/average number of conjugated drugs (Drug-to-Antibody Ratio: DAR). To this end, we have developed a range of analytical workflows employing mass spectrometry, in a regulated environment, to support pharmaceutical companies in the development and control of their mAbs and ADCs.

Characterizing Deamidation and Oxidation in Adalimumab with Low pH Peptide Mapping and Middle-Up Mass Spec Analysis.

Non-enzymatic chemical modifications such as deamidation, oxidation and disulfide bond scrambling can affect the stability and efficacy of biotherapeutic proteins. 

Quality Assistance used Biacore T200 technology to develop and validate assays allowing precise mAbs characterisation.

Our assays showed very good repeatability (intra-run precision) and intermediate precision.

Elemental impurities: how to be compliant with ICH Q3D guideline?

On January 1st 2018, all new and existing drug products will have to comply with the ICH Q3D guideline for elemental impurities (EIs). Although this guideline sets
specifications for drug products only, the risk assessment approach also involves the determination of metallic impurities in APIs and excipients.

Validation of a method for the quantification of Influenza hemagglutininin vaccine samples using surface plasmon resonance.

In vaccine production, the quantification of virus concentration is critical for the final formulation.

Comparison of multiplex assays for the quantification of cytokines

Cytokine profiling is a powerful tool to link the host immune system with disease pathogenesis...

Sanofi Pasteur has developed a new construction engineered with a genetically improved glycoprotein sequence. This glycoprotein is produced from a new recombinant cell-line developed in-house from CHO cells. The development of an improved process of production in an entirely animal-free conditions leads to a Drug Substance with a purity greater than 95%. The ability of the purification procedure to remove unwanted host cell proteins (HCPs) should be investigated thoroughly, as should the reproducibility of the process. For safety and tolerance reasons, it is necessary to guarantee that residual HCPs are reduced to an acceptable level in the medicinal product to be administered to the patient.
The objective of this study was to develop an analytical tool that allows monitoring the level of HCPs as closely as possible. Commercialized anti-CHO antibodies were compared by there recognition of HCP profiles in 1D/2D gels followed by Western blots. In order to guide the selection of the suitable antibody for the monitoring of HCP clearance from the process, we also assessed the capability of these antibodies to quantify HCPs in immunoassays. The main challenge of this study was to get rid of the dilution effect observed using different immunoassay formats.
 

In-process characterization of human mesenchymal stem cells

Cell therapy products emerge from a long process of cell culture, differentiation and purification steps where appropriate cell identity and purity monitoring are mandatory. The characterization of human MSCs used for manufacturing of cell therapy products requires the development and the validation of analytical methods to encounter biopharmaceutical requirements.

Monoclonal antibodies are among the most promising molecules for the pharmaceutical industry, but their characterization remains very challenging due to their complexity. Although mass spectrometry is now a well established technique for development work,some people are still reluctant to use it for batch release, as it is believed to be expensive and time consuming. In the present work, we present peptide mapping methodologies that can be easily used to characterize monoclonal antibodies. Tryptic peptides are usually separated by reversed-phase chromatography before UV and/or MS detection. We show that HILIC mode (Hydrophilic Interaction Liquid Chromatography) can be an interesting alternative, especially for an efficient separation of glycopeptides.

Immunogenicity assay development for biopharmaceuticals requires a case-by-case approach

Biopharmaceuticals are known to have a greater potential for inducing immune response than conventional small molecule drugs. As a significant safety and efficacy problem, immunogenicity is a major concern for regulatory agencies. Immunogenicity is usually evaluated through the detection and characterization of treatment-induced anti-drug antibodies (ADAs).

Combination of GC, UPLC, CE and MS for the characterization of monoclonal antibody glycosylation

Monoclonal antibodies are among the most promising molecules for the pharmaceutical industry, but their characterization remains very challenging due to their complexity.