Enhancing Process Rigor and Quality in RNP Manufacturing

November 29, 2023/ by Max Sellman

Webinar Recap: CRISPR Genome Editing Solutions from Discovery to Clinic

Gene-modified cell therapies have been an area of much focus for drug developers over the last decade in the cell and gene therapy arena. Innovations around CRISPR technology have made it faster and more straightforward than ever before to manufacture patient doses of gene-modified cells.

As the first generation of CRISPR medicines work their way to the clinic and towards commercial approval, drug developers and regulators are incorporating lessons learned from these initial trials into manufacturing workflows for future cures.

A critical step in the manufacturing process for CRISPR-modified cell therapies is the formation of the Cas-gRNA ribonucleoprotein (RNP) complex*. Developers and regulatory agencies alike are leveraging the importance of process control around this RNP complexation event to enhance the next generation of this medicine.

Cell therapy developers desire consistency in editing outcomes, while the FDA has issued draft guidance indicating a need to analyze the RNP complex. These processes need to be compliant with cGMP regulations to enable clinical (and eventual commercial) translation.

Deep Dive on CRISPR Data
Aldevron and Integrated DNA Technologies (IDT) have been heavily involved in the CRISPR space dating back nearly a decade. Aldevron has been at the forefront of GMP RNP manufacturing and characterization, while IDT has been an industry leader in gRNA, recently launching its GMP gRNA manufacturing services.

In a joint webinar from November 14, CRISPR Genome Editing Solutions from Discovery to Clinic, we shared details on our partnership around our latest innovations in support of GMP RNP manufacturing.

From the Aldevron side, Oat Khodthong, Ph.D., walked us through data from our three novel RNP quality control assays: free (unbound) Cas9, free gRNA, and RNP activity. These assays help to characterize GMP RNPs and can be used to ensure that the finished RNP complex meets pre-specified criteria for purity and activity.

For IDT, Garrett Rettig, Ph.D., showed us process data around IDT GMP gRNAs and functional cell-based editing results for Aldevron-manufactured RNPs. A clear conclusion stemming from this collaboration is that IDT GMP gRNA meets rigorous quality standards and that resulting RNPs produced with Aldevron nuclease are pure and functional.

During the Q&A segment, attendees were interested to learn more about the impact of gRNA:Cas9 complexation ratios on editing outcomes and RNP purity. Oat and Garrett also touched on optimization of RNP formulations and strategies to confirm production of a pure, full-length gRNA. For those working on the development of a CRISPR-based cell therapy, this webinar is a great example of how partnerships can be critical to streamline the CRISPR manufacturing workflow and critical to generate consistent and reliable data.

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