GMP (Good Manufacturing Practice) growth factors and cytokines designed for therapeutic manufacturing are a critical component in defined medias. To date, the Cell Therapy industry has accepted the term GMP for this reagent class despite the fact that there is no direct oversight by regulatory authorities. These proteins are intended to be used during further manufacturing and do not come in direct contact with the patient. In fact, Cell Therapy manufacturers need to take steps to ensure that reagents used for further manufacturing are removed before the cells can be used in the clinic. In addition, FDA-regulated, clinical grade proteins that can be directly used as therapeutics may also be described as GMP, leading to confusion of the term “GMP” within different contexts.
We recently published a blog, “GMP Proteins for Cell Therapy Manufacturing: Top 6 Things to Know,” that discusses what you should look for in a supplier. It was a popular piece with readers, so we asked the author, Tim Manning, if he would host an Ask the Expert session on the topic. Tim was kind enough to agree to answer questions from readers this week on the topic of GMP protein manufacturing.
Tim Manning is the Product Manager for Proteins at R&D Systems, a Bio-Techne brand. He has an extensive knowledge of protein production and manufacturing, quality control, and GMP guidelines. Before joining Bio-Techne, Tim received his PhD in from the University of Alabama at Birmingham and was a Postdoctoral Fellow at the University of California, San Diego.
0 0 0 Total: 0 If possible, you would prefer to use cytokines and growth factors manufactured following guidelines set by the United States Pharmacopeia (USP) and/or the European Pharmacopeia (Ph. Eur.). However, this is not always possible, and reagents made for research use only (RUO) may be the only material available. If this is … Continued
How much detail can you share about your manufacturing protocols to make sure they meet the needs of our application?
0 0 0 Total: 0 Some information can be provided via questionnaires submitted to a supplier and answered by their Quality Assurance (QA) personnel. In fact, this is often a good way to begin to ensure basic requirements can be met. Additional information is typically shared during an on-site visit or audit of the facility. … Continued
0 0 0 Total: 0 I would suggest moving to GMP as early as possible, potentially even during R&D work. This is especially the case if you are using cytokines or growth factors from a supplier that doesn’t also offer GMP for further manufacturing. Proteins are made in biological systems and are, therefore, susceptible to … Continued
0 0 0 Total: 0 Yes. Each cytokine is tested for performance using cell-based bioassays that include proliferation, induced cytokine secretion, cytotoxicity, and others. Before a cytokine is released to the public, a bioactivity specification is established based on the consistent performance of a minimum of three bulk lots. After that, each new bulk lot … Continued
0 0 0 Total: 0 The main differences come down to extensive documentation, training, oversight, and added testing. GMP proteins include Quality Assurance reviews on all GMP batch and bottling records before release. Three validation lots are made before the product is released to ensure our ability to produce consistent lots. Validated equipment and methods … Continued
I work in an academic lab and we have talked about using higher quality proteins in our research just to avoid lot variability, but it would involve a cost increase. Are there any other benefits to using GMP proteins in a research setting?
0 0 0 Total: 0 As you indicate, one of the main reasons for shifting to GMP is to ensure consistency. This comes from all of the extra analytical testing, documentation, and oversite of the manufacturing process. Protein inconsistencies can lead to negative results and missed opportunities, false positives that lead you down an incorrect … Continued
I seem to recall seeing something about the variation of animal sourced growth factors and cytokines having unexpected impacts on various cell types. Is this something you have experience with?
0 0 0 Total: 0 The vast majority of discussions we have about animal-free vs animal-sourced growth factors involve the risk of introducing pathogens. However, there are other points of variability that could be introduced by using animal-derived materials. “Animal-sourced” might mean that the natural growth factor is purified from a preparation like plasma. It … Continued
Do you have any recommendations for transitioning from research use products to cGMP produced products in terms of culture?
0 0 0 Total: 0 It’s easiest to transition if you can get GMP from the same vendor that supplies your research use only (RUO) protein. Although the manufacturing process and analytical testing is more stringent, there may be very little difference between the proteins in the vials. It is perfectly valid to ask if … Continued
We have been having some trouble with mycoplasma in our lab, we feel we are doing all the right things for good lab practices, is there any chance this be coming from our raw materials?
0 0 0 Total: 0 As you know, mycoplasma can be very hard to deal with. They are resistant to some antibiotics, may be small enough to slip through typical lab filters, and they can linger on surfaces and equipment. Some potential sources of introduction are shared cell lines between labs, contaminated reagents such as … Continued
I am interested in your recommendations for setting up secondary suppliers. What if the GMP version is hard to source? Is it ok to have a research use only as a secondary or tertiary?
0 0 0 Total: 0 It is OK to use research use only (RUO) reagents, even as a primary source when GMP for manufacturing isn’t available. If this is the case, you should still be asking the same questions of the supplier that you would if they were manufacturing using GMP for Cell Therapy. You … Continued
What kind of support is available for FDA filings. We are beginning to think about moving to GMP what should we expect from suppliers?
0 0 0 Total: 0 Some basic things to expect include a lot-specific Certificate of Analysis that will detail the identity of the protein and analytical testing that has been done to ensure the material is meeting specifications. A Certificate of Origin may be obtained that provides traceability of the materials used in manufacturing of … Continued
0 0 0 Total: 0 Unless requested differently, our GMP proteins are lyophilized in glass vials, which is typically very stable. We have environmentally controlled stability chambers that are in continuous use to ensure that there is a documented understanding of protein stability during storage. Although GMP proteins as not labeled as sterile, microbial testing … Continued
0 0 0 Total: 0 If by system, you mean cell culture vessels, this is highly dependent on the type of cell lines used and the protein being expressed. Currently these include a combination of glass, stainless steel, CellSTACK, and WAVE systems. If you mean the source, this also very dependent on the protein. In … Continued
How do you confirm the identity of your cytokines and what tests do you run for potential contaminants?
0 0 0 Total: 0 Our identity testing is performed by SDS-PAGE for molecular weight and analysis by densitometry to verify purity. We also use mass spectrometry and N-terminal sequencing of the first 10 amino acids. Endotoxin, bioburden, host cell protein, and mycoplasma testing is also done. Additional testing may be performed depending upon the protein … Continued
0 0 0 Total: 0 For the most updated list, please the GMP Page of our website. There is a link at the top of this page that will provide the latest product listing.