Development of an In Vitro Test Method to Replace an Animal-Based Potency Test for Pertactin Antigen in Multivalent...

Area of Science:

• Vaccinology
• Immunology
• Biotechnology

Background:

• Animal-based potency assays for vaccines are costly, variable, and raise ethical concerns.
• There is a need for reliable in vitro alternatives to animal testing in vaccine development.
• Pertactin (PRN) is a key antigen in DTaP-IPV vaccines, requiring robust quality control.

Purpose of the Study:

• To develop and validate an in vitro enzyme-linked immunosorbent assay (ELISA) to measure PRN antigenicity.
• To propose the developed ELISA as a replacement for animal-based potency testing of PRN in DTaP-IPV vaccines.
• To demonstrate the reliability and relevance of the in vitro assay compared to traditional methods.

Main Methods:

• Developed a PRN antigenicity ELISA using high-affinity monoclonal antibodies specific to PRN epitopes.
• Validated the ELISA for accuracy, precision, linearity, and specificity.
• Compared the performance of the ELISA with a mouse-based potency test using multiple vaccine lots.

Main Results:

• The PRN antigenicity ELISA demonstrated consistent antigenicity across vaccine lots.
• The ELISA showed higher sensitivity than the mouse-based potency test.
• The in vitro assay effectively differentiated between degraded and intact vaccine lots, outperforming the in vivo test.

Conclusions:

• The developed PRN antigenicity ELISA is a suitable in vitro replacement for the in vivo potency test for PRN in DTaP-IPV vaccines.
• Comprehensive antibody characterization, method validation, and comparison to animal testing are crucial for developing reliable in vitro assays.
• This work supports the broader strategy of replacing animal testing in vaccine quality control with validated in vitro methods.