Affinity Capture Elution (ACE) ELISA Method Development and Validation for Novel RPH-104 Drug Immunogenicity...

Area of Science:

• Bioanalytical Chemistry
• Immunology
• Pharmacology

Background:

• Growing number of therapeutic protein products necessitates robust bioanalytical methods.
• Immunogenicity evaluation is critical for therapeutic protein safety and efficacy.
• RPH-104, an Interleukin-1 (IL-1) inhibitor, is under clinical investigation for various diseases.

Purpose of the Study:

• To develop and validate a sensitive and specific bioanalytical method for immunogenicity testing of RPH-104.
• To utilize the Affinity Capture Elution (ACE) ELISA technique for this purpose.
• To ensure the method's suitability for analyzing clinical samples.

Main Methods:

• Development of an Affinity Capture Elution (ACE) ELISA assay.
• Validation of the method for key parameters: assay cut-point, sensitivity, selectivity, drug tolerance, hook effect, specificity, precision, and stability.
• Testing specificity against relevant interfering compounds (IL-1α, IL-1β, IL1-Ra).

Main Results:

• The developed ACE ELISA method demonstrated a sensitivity of 114.9 ng/mL.
• Low and high positive controls were established at 155 ng/mL and 2500 ng/mL anti-RPH-104 antibody concentrations, respectively.
• Method specificity was confirmed, and the assay was successfully applied to subject samples.

Conclusions:

• A validated ACE ELISA method for RPH-104 immunogenicity testing has been successfully developed.
• The method is sensitive, specific, and robust, meeting the requirements for bioanalytical testing.
• This assay is suitable for application in clinical studies involving RPH-104.