An analytical comparison of three immunoassay platforms for subpicomolar detection of protein biomarker GAD65

作者：Costa Olivier R^*; Verhaeghen Katrijn; Roels Sarah; Starige Geert; Ling Zhidong; Pipeleers Daniel; Gorus Frans K; Martens Geert A

来源：PLos One, 2018, 13(3): e0193670.

DOI：10.1371/journal.pone.0193670

A disproportional increase of circulating GAD65 within hours from an intraportal islet allotransplantation has been validated as biomarker of beta cell loss and poor functional outcome. More sensitive assays are, however, needed to allow detection of episodes of subtle beta cell loss during late-stage graft rejection or in the peri-onset period of type 1 diabetes. We applied the same sandwich monoclonal antibody couple reactive towards the Cand N-terminus of GAD65 on three advanced immunoassay platforms-the Cytometric Bead Array (CBA, Becton, Dickinson and Company), ElectroChemiLuminescence Immuno Assay (ECLIA, Meso Scale Discovery) and digital ELISA technology (Single Molecule Array -SIMOA, Quanterix. We then compared analytical performance (linearity, imprecision, limit of detection and functional sensitivity), correlation of results, and practicality. All evaluated techniques showed linearity up to at least 500 ng/dL (76.9 pmol/L). SIMOA achieved the lowest imprecision. The 3 platforms correlate well with each other and could all detect subpi-comolar concentrations of GAD65 in plasma, but only SIMOA and CBA could quantify down to that range. SIMOA can achieve the highest sample throughput. The three methods tested allow sensitive detection of GAD65, but SIMOA appears best suited for automated quantification of subpicomolar concentrations.

• 出版日期2018-3-8