Structural Characterization

VelaLabs is expanding its physico-chemical portfolio via the implementation of a range of diverse structure determination methods.

  

New Analytical Capabilities at VelaLabs

Recently, VelaLabs entered into the world of liquid chromatography mass spectrometry (LC-MS), gas chromatography mass spectrometry (GC-MS), size exclusion chromatography multi-angle laser light scattering (SEC-MALLS) and capillary electrophoresis (CE). Subsequent to the acquisition of our new equipment and integration of this innovative technology, we are now offering modern, up-to-date analytical methods for determining structural characteristics of peptides, proteins and small molecules.

 

 

Mass Spectrometry of Proteins: An Innovative Era in Characterizing Biological Therapeutics

Mass spectrometry (MS) is a rapidly-emerging technique to characterize protein drug therapeutics. Such proven analytical methods are now routinely used to characterize biosimilars, biopharmaceuticals & biological products during pre-clinical and clinical development as well as analysis of products during process development. The new portfolio at VelaLabs that encompasses protein mass spectrometry consists of three consecutive stages of implementation – each dedicated to support our clients in all phases of product development and before batch release.

  

Stage 1: Basic Mass Spectrometry Analysis of Your Protein

 Here we are focused on establishing and offering fundamental mass spectrometry-based analyses that include:

  • Protein identification and confirmation via gel-based and gel-free methods plus screening of different clones for biosimilar products
  • Recombinant protein intact mass measurements to, e.g., differentiate individual protein glycoforms
  • Absolute, ‘pure’ protein quantitation via spiked ‘heavy’-labelled peptides of known concentration and comparison to unknown levels in specific biological samples

  

Stage 2: Characterize and Quantitate Your Protein

For projects that require a deeper level of analysis and/or for our clients that wish to continue further from Stage 1, we can offer additional analyses such as:

  • Complete protein sequencing whereby recombinant proteins are treated with multiple proteolytic enzymes and the generated peptides are analyzed by LC-MS
  • Identification and/or localization of post-translational modifications that are important in establishing the integrity of biopharmaceuticals, e.g., assessing the degree of oxidation, de-amidation, glycosylation, phosphorylation etc. and/or protein aggregate formation
  • Global relative protein quantitation of complex mixtures via labelled and label-free approaches
  • Structural relationships to understand and identify protein-protein, protein-small molecule interactions

  

Stage 3: Tailored Approaches Dedicated to Your Requirements

 

For even more advanced projects, we are available to discuss and design specific mass-spectrometry based proteomic approaches that will involve:

  • Case-by-case discussions based on client and project requirements
  • Scientific support during process development and validation
  • Comparability studies for biosimilars to ensure that batch-to-batch variability is minimized

 

These three stages will assist in several important phases of the industrial pipelines of our Clients:

  

Discovery Phase: Non-GxP proteomic approaches will aid in: (i) clone selection; (ii) monitoring/determining in vitro proteome-wide effects; (iii) absolute quantitation of biomarkers; (iv) characterization of mutant versus wild-type protein forms via intact mass analysis and de novo sequencing; and (v) cross-validation comparison with a traditional portfolio (bioassays etc.)

  

(Pre-) clinical Studies: Here, LC-MS will provide an analytical portfolio that enables protein-based drugs measurements in complex matrices (e.g., human serum/plasma samples).  As a proof-of-principle, a simulated pharmacokinetic (PK) study on pegylated and non-pegylated granulocyte colony stimulating factor (pegG-CSF and G-CSF) and on a second biopharmaceutical, rituximab, is currently evaluated.

   

Market Authorisation: Protein mass spectrometry will be used to perform: (i) stability studies, e.g., determining the degree of protein oxidation and de-amidation of a biopharmaceutical product; and (ii) comparability studies between batches via peptide mapping and glycoform analysis. 

 

Batch Release: Via peptide mapping, LC-MS will be used to: (i) assess protein product batch-to-batch consistency; (ii) monitor/confirm recombinant amino acid sequences; and (iii) determine the degree of impurities, e.g., via host cell protein analysis. Similarly, intact protein analysis will confirm: (i) complete protein expression; and (ii) the major glycoforms and glycosylation pattern of the Protein.