BioPharm

14 BioPharm International eBook April 2019 www.biopharminternational.com Outsourcing Resources Quality Control had to do was to make analysis and reporting easier for them. In Chromeleon, we created a report template and transferred it to them directly. They would prepare a sample, then run sys- tem su itabi l it y tests on it a nd make sure they passed all assay acceptance criteria. They would then r un the test sample along with the reference standard, and could perform attribute analytics and new peak detection on the fly without having to set up these methods themselves. COST AND TIME SAVINGS BioPharm: What savings can be achieved with MAM? Rogers: To get some idea of the potential savings, consider a mol- ecule in which glycans are impor- tant, and what is involved for MAM as compared with traditional methods. Imagine that there are four relatively standard QC assays required to release that molecule: a release glycan map, identity test, charge variant test, and reduced capillary electrophoresis. Those are four separate assays that will have be qualified and validated in order to take the molecule into QC. If a new peak shows up in any one of the purity assays, that sample would need to be fractionated to isolate the new species. With MAM, you are running a digested peptide map anyway. The method's real value stems from the fact that it uses one digested sam- ple to cover all those assays and in a very analogous way, to get site- specific information for where the protein is being modified or where it is degrading. MAM data saves time and effort because it directly provides the new peak's identity. B i o P h a r m: W h a t t y p e s o f advances in model and software development were requ i red to make MAM possible initially, and what improvements have been made since then? Rogers: In the beginning of our development efforts, it was surpris- ing that we could prove proof-of- concept using Pinpoint and Sieve software. These two pieces of soft- ware were originally developed for applications that are quite different from what we were trying to do. Pinpoint, for example, was devel- oped for multiple-reaction-moni- toring (MRM)–based applications, while Sieve was designed for pro- teomics-based differential analysis. We were able to leverage both of those and show that this method could be successful. Thermo's incorporation of both attribute analytics and new peak detection into BioPharma Finder [and connecting it to Chromelon] has been a g reat step for ward. Incorporating these capabilities into a 21 CFR-compliant frame- work has allowed us to put this into a QC environment. USERS' CONSORTIUM PLANS BioPharm: What are your plans for the MAM users consortium? Roge rs: We wa nt vendors to keep innovating and improving, and we would like to work with them on developing MAM solu- tions, because that will make the method better, in the end. The consor t iu m's ma i n goa l, how- ever, is to enable the biopharma community to leverage MAM for product characterization as well as product release from QC. We are trying to improve how we do mass spec as a biopharma community, to teach people how to acquire bet- ter data and how to analyze those data more effectively using this approach. We can turn around data much quicker by leveraging this method, and can also discover potential critical quality attributes that we didn't even know were there by leveraging new peak detection. T he C onsor t iu m is foc usi ng on the free exchange of informa- tion. We currently have more than 22 0 i nd iv idu a l me mb e r s w it h more t ha n 70 compa ny mem- bers, ranging from FDA and the National Institute for Standards and Technology (NIST) from the government side, to biopharma comp a n ie s, s of t w a r e ve ndor s, instrument vendors, reagent ven- dors, and CDMOs. STANDARDIZING METHODS B i o P h a r m: W h a t i n n o v a t i v e approaches have you used recently? Rogers: In 2018, we did a Round Robin to see how well members were able to apply standardized MAM methods. Participants filled out a spreadsheet template and evaluated a reference standard, then a second version of that stan- dard that had been spiked with peptides, then a pH stress sample, and then an unknown sample. The idea was to go through all the sam- ples and look for new peaks and compare them to the reference. About half of the groups were able to pass the criteria. We see this test as the first step in teaching the industry how to apply this method in the process development lab. BioPharm: Are any companies officially using this method for QC and development, and have you published best practices on method transfer for CDMOs? Rogers: We haven't published anything yet on method transfer, although consortium members are working on projects in that topic area. So far, a couple of members have put M A M i nto t hei r QC operations. Amgen is the farthest along. FDA representatives have already visited Amgen's QC lab to see its MAM setup and to learn more about how it works. BP

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