Genetic mutation analysis at early stages of cell line development using next generation sequencing

Area of Science:

• Biopharmaceutical Development
• Molecular Biology
• Analytical Chemistry

Background:

• Biopharmaceutical companies aim to shorten development timelines to clinical proof of concept.
• Ensuring biotherapeutic material quality is crucial for clinical progression.
• Current high-throughput protein analytics assess productivity and quality, but lack genetic sequence confirmation.

Purpose of the Study:

• To explore the utility of Next-Generation Sequencing (NGS) for genetic sequence confirmation in early-stage cell line development.
• To address limitations of traditional mutation analysis techniques (e.g., mass spectrometry) in terms of sample number and turnaround time.
• To integrate a robust NGS method into the biotherapeutic development workflow.

Main Methods:

• Utilized amplicon sequencing, a Next-Generation Sequencing (NGS) technique.
• Developed and incorporated an NGS method into the cell line development workflow.
• Applied the NGS method alongside other high-throughput protein analytical assays.

Main Results:

• The NGS method was successfully integrated into the cell line development workflow.
• The NGS method identified at least one Chinese hamster ovary (CHO) clone expressing a variant biotherapeutic form in each of the four clinical programs evaluated.
• The method proved robust, rapid, sensitive, and amenable to multiplexing.

Conclusions:

• NGS, specifically amplicon sequencing, is a valuable tool for genetic sequence confirmation in biotherapeutic development.
• This sequence confirmation method is essential for accelerating the time to clinical proof of concept.
• Implementing NGS guards against delays caused by sequence mutations, ensuring safer and more efficient biotherapeutic development.