This case study demonstrates how Sekbio's optimized antibody pairs enabled a leading IVD manufacturer to significantly improve the sensitivity and reliability of their cardiac biomarker CLIA assay.
Challenge Background
The client, a major diagnostic company, was developing a high-sensitivity cardiac troponin I (cTnI) assay but struggled with:
- Inconsistent results at low concentrations
- High background signal
- Limited detection range
- Variable performance between lots
Requirements
Target specifications:
- Limit of detection (LOD): < 2 pg/mL
- Linear range: 2-50,000 pg/mL
- Intra-assay CV: < 5%
- Inter-assay CV: < 8%
- Shelf life: 18 months
Solution Approach
1. Antibody Selection
We screened over 200 antibody pairs targeting different epitopes of cTnI, identifying optimal capture and detection combinations.
2. Epitope Mapping
Utilized peptide array technology to map epitopes, ensuring minimal interference from other cardiac proteins.
3. Conjugation Optimization
Developed proprietary HRP conjugation protocol maximizing activity while maintaining stability.
4. Buffer Optimization
Designed assay buffer minimizing non-specific binding and maximizing signal-to-noise ratio.
Results
- LOD: 1.5 pg/mL (exceeding target)
- Linear range: 1.5-100,000 pg/mL (expanded range)
- Intra-assay CV: 2.3% (excellent)
- Inter-assay CV: 5.1% (improved)
- Shelf life: 24 months at 2-8°C
Implementation Impact
Post-implementation benefits:
- Reduced reagent costs by 30%
- Improved assay throughput by 25%
- Decreased time-to-market by 3 months
- Enhanced customer satisfaction
- Strengthened market position
Lessons Learned
- Epitope selection is critical for assay performance
- Antibody quality directly impacts clinical utility
- Early-stage collaboration saves development time
- Comprehensive characterization prevents late-stage issues