The analytical characterisation of monoclonal antibodies is complex and requires a wide range of techniques, due to high heterogeneity of these proteins. This heterogeneity is due to post-translational modifications such as glycosylation, C-terminal lysine clipping, oxidation, deamidation, etc. Charge variant analysis is required to detect and quantify some of these modifications for characterisation purposes, for batch release or during stability studies.

Quality Assistance is equiped with state-of-the-art technologies for charge variant analysis, including ion-exchange chromatography and imaging capillary isoelectrofocusing (icIEF). These techniques can be run in a full GMP compliant environment and can therefore be used at the different stages of the development of a therapeutic monoclonal antibody.

During this live webinar, we will outline the different analytical workflows that can be used for charge variant analysis (ion-exchange chromatography with salt or pH gradients, MS-compatible approaches, 2D-LC/MS, and icIEF) and present several case studies.

Download the slides of 45 minutes live webinar to learn how Quality Assistance has developed different analytical workflows that can be used for charge variant analysis (ion-exchange chromatography with salt or pH gradients, MS-compatible approaches, 2D-LC/MS, and icIEF) and how you could benefit from this expertise.

Access the slides of this webinar highlighting an analytical method we have developed in our labs for the absolute quantification of proteins by ICP/MS. This method holds the potential to become the new gold standard for protein quantification, in place of amino acid analysis.

Mass spectrometry is a valuable tool for the development of biotherapeutics. This technique can be used in a GxP environment at every step of process development and for product optimisation.

Follow R&D Director, Arnaud Delobel, as he presents you our Mass Spectrometry installations, the analytical services we provide and our current R&D projects.

Access the slides of the webcast now to learn more about the services we provide and the reasons why you should partner with us.

Absolute quantification of proteins (i.e. quantification without the need for a reference standard) is a key analysis for the qualification of reference standards or to determine the extinction coefficient of a protein. It is usually performed using amino acid analysis, but this technique shows limited accuracy and repeatability.

Quality Assistance has developed an innovative method based on the quantification of sulphur by isotope dilution ICP/MS. Unprecedented precision and accuracy can be obtained, which provides a reliable quantification of proteins and determination of extinction coefficients.

Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) is a powerful technique for the advanced structural characterisation of proteins and the study of protein-protein interactions.This technique has been used for several decades, mostly in academic labs. Thanks to advances in automation, liquid chromatography and mass spectrometry technologies, this technique can now be used to get reproducible and reliable results in characterisation studies of biotherapeutics. It is complementary to other structural biology techniques such as NMR and X-ray cristallography. Its main advantage is to work in nearly physiological conditions and to require relatively limited amounts of sample.

Access this 45 minutes webinar to learn how HDX-MS can be applied throughout the development of your product to get structural information that can hardly be obtained with other techniques. After a detailed description of the technique and its applications, case studies will be presented in the context of comparability and epitope mapping studies. 
 

Applications of Multiple-heartcut 2D-LC/MS at Quality Assistance:

• Characterisation of monoclonal antibodies

• Characterisation of ADCs : 
  Hydrophobic Interaction Chromatography (HIC) is a method of choice for the characterisation of ADCs. Due to high salt content in mobile phase, it is impossible to hyphenate the chromatography to MS for peak identification. Fraction collection and desalting is time-consuming and is not easily amenable to low concentration species (due to dilution effects)

• Quantification of free drugs in ADCs:
  Quantification of free drug in ADCs is critical for batch release and during stability studies. Cytotoxic payloads are extremely toxic and low limit of quantifications should be achieved. MS is often required to reach low LOQ, and the protein has to be removed (offline or online) before the analysis. 2D-LC/MS is a way to meet all those criteria

• Analysis of in-process samples (work in progress)

Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) is an innovative tool for the characterisation of biotherapeutics. Besides the study of higher-order structures, HDX-MS can also be used for epitope mapping studies in order to determine to which region of the target a monoclonal antibody binds. HDX-MS is a good alternative to techniques such as X-ray diffraction or NMR.

Glycosylation is critical for the safety and efficacy of biotherapeutics.

Orthogonal methods have to be applied for characterisation studies and QC testing. LC/MS is a valuable tool for glycosylation analyses. All these methods can be applied in a regulated environment.