Addressing Regulatory Requirements for CRISPR sgRNA Purity with Orthogonal Chromatography

Learn about a novel dual-purification method for CRISPR sgRNA. Discover how this approach achieves over 80% purity and meets new regulatory standards.

Register Free: https://www.biopharminternational.com/bp_w/orthogonal-chromatography

Event Overview:

Join us for a webinar with Agilent’s experts to explore a novel purification method for CRISPR single guide RNAs (sgRNA). This approach, proven under GMP, achieves over 80% full-length purity confirmed by high-resolution HPLC methods and meets new regulatory standards.

Some CDMOs claim to achieve higher than 80% or even 90% purity of sgRNA, but it’s important to distinguish the methods used to assess purity and their resolution power. Without the comparability of resolution power the purity from one vendor to another may be widely varying (greater than +/- 10% purity).

Recent regulatory guidelines from the FDA’s Center for Biologics Evaluation and Research (CBER) emphasizes the need to achieve 80% purity levels for the full-length sgRNA product.

To address the new requirements, Agilent has developed an orthogonal chromatography approach using 2 modes of separation. The approach features Agilent’s PLRP-S purification resin that complements a strong anion exhange purification. Varying-length gRNAs up to 120 nucleotides (nt) in length have been able to achieve over 80% purity using this dual-purification strategy. Translation of the technique into GMP production has also been achieved.

Additionally, there will be a discussion of a novel MS-guided in-line PAT (process analytical technology) sequence verification during solid-phase synthesis that confirms the sequence for all nucleotide types, including chemical modifications. In the first sentence, should “single guide RNAs” be “single guide RNA” to match the abbreviation that follows?

Key Learning Objective:

• FDA CBER guidance is for greater than 80% purity levels for full-length sgRNA products.
• How resolution power of the analytical chromatography purity methods allows for comparison of quality
• How using Agilent’s optimized oligonucleotide synthesis approach and its new orthogonal chromatography approach can reliably achieve 80% purity.
• MS-guided sequence verification (MSV) ensuring accurate and reliable results.

Who Should Attend:

• Gene editing innovators in pharma and biopharma
• Analytical development, manufacturing, process development, and quality control experts and leaders

Speakers:

Amanda Haas
Business Development Manager, Nucleic Acid Solutions
Agilent Technologies

Amanda Haas is a business development manager at Agilent Technologies with over 19 years of experience in life sciences and biotechnology. She has been with Agilent for 3 years, growing gene-editing therapeutic services using market research, voice of the customer, and collaboration with the development team. Amanda holds a BS degree in chemistry from the University of Miami. She previously worked at Horizon Discovery, Dharmacon, and Thermo Fisher Scientific.

Joe Guiles
Head of Development, Nucleic Acid Solutions
Agilent Technologies

Register Free: https://www.biopharminternational.com/bp_w/orthogonal-chromatography