Selection and Characterization of Cell Line-Virus Pairs for Sensitive Viral Detection Assays in Biopharmaceutical...

Area of Science:

• Biopharmaceutical Manufacturing
• Virology
• Cell Biology

Background:

• Viral safety is paramount in biopharmaceutical production, necessitating robust detection methods for adventitious agents.
• In vitro viral detection assays rely on specific cell line-virus interactions to identify contaminants.
• Optimizing these interactions is key to ensuring assay sensitivity and reliability.

Purpose of the Study:

• To systematically evaluate cell line-virus combinations for optimal in vitro viral detection.
• To identify the most sensitive and reliable models for detecting adventitious agents in biopharmaceuticals.
• To establish a framework for selecting appropriate detection systems in quality control.

Main Methods:

• Analysis of three cell lines (Vero, MRC-5, BHK-21 [C-13]) against four model viruses (Reovirus type 3, Adenovirus type 5, Human parainfluenza virus type 3, Herpes simplex virus).
• Evaluation of cytopathic effect (CPE) kinetics, morphology, and detection sensitivity (Limit of Quantification - LOQ).
• Systematic comparison of infection dynamics and CPE characteristics for each combination.

Main Results:

• All tested systems showed high analytical sensitivity with LOQ as low as 0.01 TCID50/mL.
• Significant variations in infection dynamics and CPE morphology were observed across different cell line-virus pairs.
• BHK-21 [C-13]/Reovirus type 3, Vero/Adenovirus type 5, and MRC-5/Human parainfluenza virus type 3 and Herpes simplex virus were identified as optimal combinations.

Conclusions:

• Assay performance is influenced by both sensitivity and the specific kinetics and morphology of viral infection.
• Rational selection of cell line-virus pairs is essential for developing effective visual screening assays.
• The identified optimal pairs provide a foundation for routine viral detection in biopharmaceutical quality control.