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The CEPharm Troubleshooting Parties:

At CEPharm 2019, we used the troubleshooting workshop to focus on a more concerted effort to “get rid of” two issues that have been following/nagging the CEPharm community for some time and that we want to get out of the way in order to be able to focus on other things. We took the troubleshooting workshop to set up two inter-company collaborations, one on CE-SDS ghost peaks and baseline dips, and one on amino caproic acid reagent variability. More info here.

CE-SDS Ghost Peaks and Wobbly Baselines

The ghost-peaks and baseline dips have been filling many troubleshooting sessions in the past years and most of us recognize these. Therefore we think it is high time that we collect and document all we as CE community know, suspect or guess about the ghost peaks and make this knowledge accessible to all. First, we would like to publish the gathered “tribal” knowledge in a review paper. If it turns out that there are still remaining ghost peaks that escape all our knowledge and experience, we will form hypotheses on the potential root causes and perform collaborative experiments to challenge the hypotheses. Results should then be collected, analyzed, presented at CEPharm, and ultimately published.

Party-leader: Kristin Schultz Kuszak, AstraZeneca
Support: Tim Blanc, Eli Lilly and Cari Sänger - van de Griend, Kantisto

eACA Reagent Variability

Many of us have embraced CZE for protein analysis. The well-known application published by Pfizer is being used by many companies. During the past troubleshooting sessions, several problems attributed to the eACA quality were shown. Different users have done experiments, but we still don’t know exactly what causes the issues and what quality requirements we need to put on eACA buffers. First, information from different users will be collected about supplier, batches, capillaries, detectors etc. and look for trends in these data. As a next step, hypotheses will be formed on root causes and experiments will need to be performed by multiple people to confirm or decline the formed hypotheses and define practical solutions. Ultimately, we want to gather the knowledge obtained, share this at CEPharm, and publish it.

Party-leader: Rebecca Wiesner, Technical University Braunschweig
Support: Hermann Wätzig, Technical University Braunschweig and Cari Sänger - van de Griend, Kantisto

Data collection

In this multi-company (and vendor, regulatory agencies and institutions) study, we want to collect the method-related data to the general improvement of pharmaceutical analysis for patient safety and efficacy. In addition, this is also of the contributing laboratories’ interests, as it will reduce the amount of failure rates and reanalysis. We do not collect analyte-specific data. Several companies have already contributed. In order to safeguard the contributors, an independent and neutral person will anonymize all the data before it is shared with the group.

We organize these troubleshooting working parties specifically and purposefully as CASSS, which is a not-for-profit organization. The mission and vision are copied here:

  • CASSS is an agile, non-profit scientific organization whose strength is in bringing together professionals from industry, academia and regulatory agencies to solve scientific and technical problems in order to advance the development of biopharmaceuticals.  CASSS enables regulatory capacity building, knowledge-sharing and global access by leveraging strategic partnerships, scientific expertise and a diversity of views in all of our activities.
  • CASSS exists today to enable a global community of industry, academic and regulatory professionals to work together to resolve scientific challenges in the field of biopharmaceutical development and regulation.
  • CASSS’ community includes process, analytical, regulatory and other related scientists in the field of biopharmaceutical development and regulation.

CASSS CORE VALUES: Integrity, Community, Purpose, Collaboration, Distinction

We all know it and we all hate it: the dreaded “ghost peak” in reduced CE-SDS (see below).  He is a devious one that appears randomly and disappears before you can understand the root cause.  Let’s put this peak to sleep!  Here is what we know about him:

  1. He appears between the two non-reducible species peaks (~24 minute retention time  with the 20 cm meter separation) 
  2. Sample buffers with pHs ranging from 6.5 to 9.0 produce this peak
  3. His magnitude can vary: 0.2% up to 2.0% time correct area
  4. Re-injection from the same vial will replicate this peak (i.e. it does not go away with time)
  5. Only re-prepping the sample will make it disappear
  6. Heat sources and vials have been the targets of investigations in the past
    • These are contributing factors, but not the prime cause
  7. Some molecules produce this peak easier than others
  8. Changes in reducing agent concentration can mitigate the peak

We would like to see if there is a molecule characteristic, which when combined with the faulty heat sources/vials could manifest in the ghost peak. Please leave your comments through the contact form as to how you and your company have mitigated this peak and/or if you have discovered any possible links between structure function characteristics and the manifestation of this peak.

 

More information on the CEPharm CE-SDS Troubleshooting Parties and data collection.

We organize these troubleshooting working parties specifically and purposefully as CASSS, which is a not-for-profit organization.