GEN: Let’s begin by finding out more about Entegris. Your organization serves the microelectronics, industrial, and life sciences industries. Here’s more info on rapid prototype (www.easyfie.com) look into our web site. What technologies do you carry to the life sciences table?
Dr. Chen: Entegris was founded in 1966 and operates out of our headquarters in Billerica, MA. Now we have a worldwide footprint in North America, Europe, and Asia with 5,800 employees worldwide. We focus on providing contamination management, critical material handling, and superior process supplies used in the semiconductor device fabrication course of. Our traditional customers include semiconductor device manufacturers, OEMs that provide tools to semiconductor system manufacturers, gas and chemical manufacturing companies, main wafer grower companies, and manufacturers of high-precision electronics. Intel, Samsung, and TSMC are a couple of of our clients. As a technology-pushed firm, we have now 2,520 active patents and an extra 1,050 pending ones.
Entegris officially branched into the life sciences industry about two years in the past. We see that our core technologies will help deal with the unmet wants of the life sciences enterprise. We deliver fifty five years of experience in filtration and purification, fluid administration, and manufacturing excellence to the life sciences desk. Our single-use assemblies are utilized by main COVID-19 vaccine suppliers as a consequence of their extraordinarily low extractable and leachable profile, gamma compatibility, wide working temperature window, and high chemical compatibility.
Our experience in filtration and purification to ppt (parts per trillion) levels positions us nicely for life sciences filtration functions. Our global manufacturing footprint, specialties in ultraclean components and materials, rapid prototyping, and customization give us unique benefits in addressing unmet life sciences business wants.
GEN: Entegris has been working with FUJIFILM Diosynth Biotechnologies (FDB) on a mission entitled “A material characterization study for AAV process growth and tool selection.” Tell us slightly more about the main focus of the mission and what brought the two corporations together.
Dr. Chen: Our clients have used Aramus fluoropolymer (FP) single-use luggage and assemblies for all times sciences functions corresponding to COVID-19 vaccine manufacturing, cell and gene therapy, bulk drug substance (BDS) storage, and high-cell-density banking. With elevated adoption of fluoropolymer materials into life sciences functions, we wish to supply our prospects with more software-particular info on our merchandise. We now have information accessible for extractables and leachables, mechanical properties, working temperature range, permeability, chemical compatibility, gamma compatibility, and many others.
However, we cannot discover a lot data in the literature in regards to the adsorption properties of fluoropolymers in contrast with completely different materials used in cell and gene therapy applications. With the discharge of the FDA’s new steerage on chemistry, manufacturing, and control (CMC) for human gene therapy investigational new drug applications (INDs) in 2020, choosing optimal materials to ensure early-phase regulatory compliance has once again been introduced into the highlight.
We needed to study the adsorptive behavior of viral vectors typically used for gene therapy using Aramus fluoropolymer materials in an effort to assist biomanufacturers reduce product losses throughout manufacturing, simplify in-course of procedures for titer determination, and scale back formulation threat with inaccurate titers. FDB is a contract and improvement manufacturing organization (CDMO) in a position to offer finish-to-end providers which include the development of cGMP-compliant processes for manufacturing viral vectors and the analytical capabilities wanted to assist batch-launch testing and in-depth product characterization. The company’s deep understanding of virology and its experience in viral vector manufacturing and evaluation is complementary to our expertise in single-use applied sciences and solutions.
FDB, which has developed a triple transfection-based platform for the manufacture of AAV viral vector-primarily based therapeutics, is always striving for the best high quality and greatest yield doable. As FDB developed this platform course of, it also identified the “hold” steps within the downstream course of as areas that could use better management. The collaboration between Entegris and FDB aimed to search out options to these challenges.
GEN: Briefly provide the key details about what was truly completed through the examine. What was the ultimate end result?
Dr. Chen: To fulfill Entegris’ venture wants, the FDB group produced high-titer AAV viral vectors consultant of fabric in the bulk drug substance stage of the manufacturing course of. To visualize the product loss throughout storage, FDB evaluated multiple titer methods for both genome copy number and whole viral particle quantity. As a result of variabilities in these assays, FDB chose an SE-HPLC method for particle titer determination and evaluated the particles recovered from samples taken over the time course with particle titer determination of the input materials. The SE-HPLC method permits a extra fast, sensitive, and reproducible measurement of AAV viral particle titer than with prevalent qPCR methodologies.
Solutions containing accurately decided amounts of AAV particles have been stored in single-use polyethylene (PE), ethylene vinyl acetate (EVA), and FP baggage at room temperature. The baggage had been sampled at time intervals as much as four days, and AAV concentrations have been immediately decided by SE-HPLC and in comparison with the starting amount.
The outcomes demonstrated that while the HPLC peak area associated to AAV concentration in PE baggage decreased by greater than 15% over 4 days in comparison with that of the beginning AAV concentration, the AAV concentration in FP bags confirmed minimal to no lower for as much as four days at room temperature, which performed much like EVA luggage for this AAV serotype.
GEN: Why was the Aramus 2D single-use meeting capable of exhibit the performance it demonstrated by the study?
Dr. Chen: The Aramus 2D single-use meeting is manufactured from single-resin, additive-free, and gamma-stable fluoropolymer film with low surface power, which makes it extra non-adsorptive and appropriate for this software.
GEN: What conclusions might be drawn from the Entegris-FDB study?
Dr. Chen: There have been four essential conclusions from the study:
1. Single-use FP luggage reveal suitability for storage of AAV- containing options in the course of the manufacturing course of because of their minimal adsorption of AAV particles, in contrast to extra commonly used PE bags. Single-use FP bags also demonstrated on-par adsorption efficiency with EVA bags together with the conical-tube management in this examine. Choosing fluoropolymer materials can help avoid or decrease product loss throughout AAV manufacturing processes and cut back the complexity of quantification and formulation risks throughout closing fill and end.
2. The collaboration between Entegris and FDB has highlighted the need for a more rapid, delicate, accurate, and precise technique to determine AAV focus throughout manufacturing processes.
3. During this collaboration, FDB has developed a extra fast, sensitive, and precise SE-HPLC methodology to quantify particle titers for his or her viral vector platform, which performs higher than the qPCR method.
4. There may be a need for the event of more amenable in-process analytical methods to support the rapid growth and scale-up of AAV manufacturing processes while maintaining compliance with regulatory necessities.