From polyclonal to monoclonal: de novo sequencing of goat antibodies for a standardized ApoA-I immunoturbidimetric assay
peer-reviewed · Journal of Biological Engineering · 2026
| Date | 2026-05-12 |
| Type | peer-reviewed |
| Venue | Journal of Biological Engineering |
| Publisher | Springer Science and Business Media LLC |
| Contribution | downstream-application |
| DOI | 10.1186/s13036-026-00697-y |
| Citations (OpenAlex) | 0 |
Abstract
Antibodies serve as essential recognition elements in in vitro diagnostics (IVD), yet reliance on polyclonal antibodies (pAbs) introduces significant challenges related to batch-to-batch consistency. To address this, we employed a de novo sequencing platform, which identified 15 candidate monoclonal antibody (mAb) sequences. From these, four distinct mAb mixtures were formulated and evaluated. Using an immunoturbidimetric assay to quantify apolipoprotein A-I (ApoA-I) in 105 human serum samples, the performance of these mixtures was compared to that of a conventional pAb reagent. The four-mAb mixture demonstrated excellent correlation with the pAb assay (Passing-Bablok: y = 1.0536x-0.0748 g/L; r 2 = 0.994). This optimized cocktail successfully replaced the pAb reagent, matching or exceeding its performance in key analytical parameters including linearity, sensitivity, and precision. Our work thus establishes a practical pathway for converting heterogeneous pAbs into defined, recombinant antibody reagents suitable for standardized IVD applications.
Methods and tools
- Goat ApoA-I antibody de novo sequencing: Replaces a polyclonal reagent with defined monoclonals by de novo sequencing goat antibodies: 15 candidate sequences were recovered, four mixtures formulated, and the four-antibody mixture benchmarked against the polyclonal in an ApoA-I immunoturbidimetric assay over 105 human sera.