Streamlining Charge Variant Analysis with MauriceFlex
Posted August 28, 2024
"Adding fractionation capabilities can really help in simplifying the workflow to assess peak identity, especially if paired with state-of-the-art mass spectrometry techniques. On top of this, the experience of working with the Bio-Techne family has been a pleasure so far and they helped us throughout our first work, beyond the training aspect of the installation."
- Dr. Sara Carillo, Bioanalytical Research Lead, NIBRT
Listen to Dr. Carillo on how she is using the MauriceFlex™ platform for charge variant characterization using mass spec.
- Can you provide us a short summary of your professional background and when and why you joined NIBRT?
I have a chemistry background and, before joining NIBRT (National Institute for Bioprocessing Research and Training), my interest was on macromolecular structures present on Gram-negative bacteria and how they were interacting with the host immune system. Structural analysis by mass spectrometry was a critical part of my work and when I joined NIBRT, I had the opportunity to apply my learnings to biopharmaceuticals. As such, exploiting every available tool to simplify product intrinsic heterogeneity is pivotal to deep and confident characterization of these molecules through mass spectrometry. For this reason, our group in NIBRT has a strong focus on separation techniques coupled with mass spectrometry detection.
- What is the mission of NIBRT, and how does your role and the work you do at NIBRT contribute to that mission?
NIBRT’s mission is to address the important needs of the biopharmaceutical manufacturing industry in Ireland and internationally, and ultimately to help deliver biologic-based medicines to people who need them. In particular, the work of the Characterization and Comparability Laboratory lead by Prof. Jonathan Bones is focused on developing analytical solutions to complex scientific problems that deliver utility and impact and facilitate advancements in biopharmaceutical manufacturing. This means that we need to aid the advancement of the bioprocess pipeline from drug discovery and early drug development down to quality control and release tests. Each phase of the bioprocess pipeline has its own unique challenges and the increased complexity of the biopharmaceutical landscape requires our methodologies to be continuously improved to return more and more accurate data and to fully exploit the analytical technologies advancement available on the market.
- Tell us about the decision to bring MauriceFlex™ into NIBRT.
As already mentioned, the possibility to simplify the complex proteoform mixture is a critical need to assess product profiles during all phases of bioproduction or, for example, to optimize the profile of biosimilar candidates. The gold standard for charge variant profiling is icIEF and with the MauriceFlex we are now able to couple this separation technique with mass spectrometry for direct identification of the different variants. This brings a new range of possibilities to further develop our workflows and further understand complex samples such as bispecific antibodies, antibody drug conjugates or gene therapy related products, such as AAV derived viral proteins.
- Describe the experience of getting the MauriceFlex up and running. How long did it take?
MauriceFlex was installed in NIBRT over 2 days and training arranged immediately afterward. During the training it was already possible to analyze some of our samples of interest so it was a pretty smooth process before being ready to develop and optimize our workflows.
- Has MauriceFlex replaced any other methods?
MauriceFlex was the first instrument that allowed us to perform icIEF separation in our laboratories, expanding our ability to determine the presence and quantify acid and basic species in our samples. We still use capillary electrophoresis and ion exchange chromatography to support and complement our data or according to the specific project needs. Compared to direct hyphenation with mass spectrometry, MauriceFlex will allow us to retain the single components for future analysis and site-specific determination of the modifications causing the pI shift.
- What methods does the MauriceFlex complement, and why? Tell us about integrating the MauriceFlex with different downstream mass spec analysis. How does it compare to other methods?
As already mentioned, MauriceFlex can nicely complement charge variant information coming from other separation techniques. The ability to fractionate the sample for future use gives the analyst the possibility to investigate peak identity further if coupled with subunits analysis or peptide mapping experiments. Moreover, the flexibility to move from an analytical to a fractionation setup provides good transferability across a QC environment and R&D, allowing to easily replicate an experiment if an unexpected peak is detected and to quickly verify its identity.
- Tell us about any posters or publications that you are working on.
We started to enroll the MauriceFlex on a comparability study across infliximab biosimilars. Although many studies have compared the originator and biosimilar products, charge variant profiles apparently similar are masking the presence of proteoforms with different pI values. If we can streamline a robust, quick and more accurate identification of charge variant peaks, we feel we can massively improve biosimilar production, both during development and clone screening, but also during the approval process.
- How do you see MauriceFlex’s role in your future biomolecular characterization projects?
We are surely observing a shift in the current landscape, with methods for monoclonal antibodies becoming platform and requiring increased robustness and shorter turnaround times between analytical teams and manufacturing. At the same time, the more complex formats and gene therapy related products are still hard to decipher and constitute an actual critical challenge to bring these products to the next level. It will be fundamental to have tools able to help the analytical scientists to understand every aspect of the product and icIEF technologies like the MauriceFlex are putting us on the right track to solve some of these puzzles. Moreover, as more biopharmaceuticals will come off patent in the coming years, the ability to not only compare but to fully characterize charge variant profiles will be important to develop more biosimilars. The final result will be that life-changing drugs will be more affordable and more available, improving patients experience.
- What advice do you have for customers considering bringing MauriceFlex into their biomolecular characterization workflow?
I believe icIEF capabilities are already present in the majority of the laboratories in the biopharmaceutical sector. Adding fractionation capabilities can really help in simplifying the workflow to assess peak identity, especially if paired with state-of-the-art mass spectrometry techniques. On top of this, the experience of working with the Bio-Techne family has been a pleasure so far and they helped us throughout our first work, beyond the training aspect of the installation.
- What advice would you give to students who are interested in pursuing a career in a role that supports the manufacturing and analysis of biologic medicines?
Developing methodologies that support biopharmaceuticals discovery and production allows you to see an immediate impact of your research on the real world and on the quality and the efficiency of how these drugs are produced. During conferences we meet analysts working in biopharma companies that are following our work and try to establish what we developed in NIBRT in their facilities. The workflows end up improving the analysis of a real product that is helping patients to go through the struggles posed by a disease. This is priceless and rewarding.