Project Examples – Optimization of an ELISA Method to Increase Assay Sensitivity

Objective: To improve the sensitivity of an ELISA that is specific for a TGF-B gene family member approximately 100 fold.

Performance Specification: Analytical sensitivity < 10 pg/mL

Proposed Timeline: 6 Weeks

Description: In Phase 1, IBA evaluated the use of competitive and non-competitive immunoassay formats employing different enzymatic detection systems to achieve the required sensitivity.  The client's ELISA was established in house and the key assay parameters (reagent concentrations, volumes, assay incubation times and temperatures) were optimized. 

In Phase 2, additional antibodies were evaluated in competitive and non-competitive formats for the required sensitivity. Antibodies that were suitable were optimized in the appropriate immunoassay format.

Results: Antibodies were identified that were suitable for the detection of the analyte in either sandwich or competitive ELISA formats.  We identified and optimized an antibody pair that when used in a sandwich immunoassay format achieved an analytical sensitivity of 4 to 8 pg/mL.

Project Completion: 6 Weeks