Overcoming Biopharmaceutical Interferents for Quantitation of Host Cell DNA Using an Automated, High-Throughput...

Area of Science:

• Biopharmaceutical Analysis
• Analytical Chemistry
• Vaccine Development

Background:

• Biopharmaceutical development requires robust assays for characterizing complex products.
• Residual host cell DNA and protein are critical impurities impacting product safety and efficacy.
• Existing methods may struggle with complex matrices and diverse biopharmaceutical samples.

Purpose of the Study:

• To develop a novel, high-throughput assay for quantifying residual host cell DNA in vaccine drug substances.
• To address challenges posed by sample matrix interferences, such as RNA and proteins.
• To establish a cost-effective and rapid analytical method for biopharmaceutical process monitoring.

Main Methods:

• Utilized a commercially available fluorescent dye (Picogreen) in a 96-well plate format.
• Implemented an enzymatic digestion pre-treatment to remove interfering substances like RNA and proteins.
• Incorporated deoxycholate to disrupt sample matrix interactions for selective DNA quantitation.

Main Results:

• Successfully developed and validated a high-throughput host cell DNA quantitation assay.
• Demonstrated the assay's ability to overcome interferences from RNA, proteins, and complex sample matrices.
• Evaluated and confirmed critical analytical figures of merit, including precision and spike recovery.

Conclusions:

• The novel assay provides accurate and selective DNA quantitation for diverse biopharmaceutical samples.
• The method is suitable for both upstream and downstream process analysis in biologics and vaccine manufacturing.
• This automated, high-throughput approach enhances efficiency and reliability in biopharmaceutical quality control.