Project Examples – Development of Multiplexed Immunoassays to Quanticate Human IgG Antibodies to a Polyvalent Viral Vaccine
Objective: To develop and optimize a multiplexed immunoassay to quantitate human serum IgG antibodies to four individual serotypes of virus-like particles (VLPs).
Performance Specifications: Analytical sensitivity and specificity equal to or greater than the client’s single analyte ELISAs.
Correlation coefficient (R2) > 0.800 in a comparison of patient sample anti-VLP serotype-specific IgGs determined in multiplexed and single analyte ELISAs.
Proposed Timeline: 12 Weeks
Description: IBA employed the BioPlex™ multianalyte detection system and optimized the conjugation of specific VLPs to four separate bead sets to capture antigen-specific patient IgG antibodies. Patient antibodies were detected by sequential addition of commercially available anti-human IgG immunoreagent conjugated to biotin and strepavidin conjugated to phycoerythrin. Antibody quantitation was determined by reference to a multiplexed human serum IgG calibrator containing IgG reactivity to each of the four VLP serotypes. Anti-IgG reactivities in the multiplexed calibrator serum were value assigned based on reference standards.
Results: Multiplexed assay sensitivities and specificities achieved were equal to or better than the single analyte ELISA sensitivities and specificities.
- R2 > 0.810 for all four multiplexed assays.
- Total assay time was less than 3 hours.
- Patient serum required for the multiplexed assay was 1 to 2 μL.
Project Completion: 8 Weeks
|
