Specific detection of tetanus toxoid using an aptamer-based matrix

Area of Science:

• Biotechnology
• Immunology
• Analytical Chemistry

Background:

• Biological production of therapeutic proteins necessitates stringent batch monitoring.
• Current animal-based risk assessments for biologicals are ethically and economically unfavorable.
• In vitro methods are being developed to replace animal testing, with enzyme-linked immunosorbent assay (ELISA) as a standard for tetanus toxoid integrity.

Purpose of the Study:

• To develop a novel detection method for tetanus toxoid using RNA aptamers.
• To evaluate the performance of aptamer-based assays compared to traditional antibody-based ELISA.
• To assess the stability and robustness of aptamer-based detection under various stress conditions.

Main Methods:

• Identification of RNA aptamers specific for tetanus toxoid using displacement assays.
• Development of a sandwich aptamer-linked immobilized sorbent assay (ALISA) using selected aptamer pairs.
• Stress testing (thermal, mechanical, freeze-thaw) of adjuvanted tetanus toxoid and analysis by ALISA and ELISA.

Main Results:

• RNA aptamers demonstrated high specificity and selectivity for tetanus toxoid detection.
• The ALISA format achieved sensitivity comparable to conventional ELISA.
• Aptamer-based detection showed similar results to ELISA when assessing antigenicity loss under stress conditions.

Conclusions:

• RNA aptamers can effectively replace antibodies in detection assays for tetanus toxoid.
• Aptamer-based assays offer a more stable and reproducible alternative to ELISA.
• This approach facilitates the development of robust and ethical tools for biological product quality control.