An Optically Silent Epoxysilane Chemistry for Paper-Based ABO Blood Typing

Safe transfusion depends on rapid, accurate ABO typing, yet reference methods require centrifuges and instrumentation unavailable at the point of need. Here, a paper-based ABO-RhD typing device built on a covalent, optically silent surface chemistry is reported. Aminosilane (APTES) and epoxysilane (GPTMS) functionalisation of Whatman cellulose was compared by water contact angle, energy-dispersive X-ray spectroscopy (SEM-EDS) and infrared spectroscopy (FT-IR) across two paper grades, three silane concentrations (5, 10 and 20% v/v) and three reaction times (1–6 h). APTES produced a strongly hydrophobic layer that impeded aqueous wicking, and its glutaraldehyde activation generated a red-brick chromophore incompatible with a red-channel readout. GPTMS coupled antibodies in a single mild step, without a crosslinker or visible chromophore, while preserving wicking. GPTMS (10% v/v, 3 h, Whatman No. 4) with a six-cycle 100 µL saline wash was selected; antibodies were immobilised in a four-zone layout (anti-A, anti-B, anti-D and control) within a 3D-printed two-compartment housing that traps agglutinated cells while free cells wash through. On 80 EDTA clinical blood samples (20 each of groups A, B, AB and O) the device classified every sample correctly (accuracy 100%; 95% confidence interval 95.4–100%), with visual and instrumented reads in full agreement. All samples were RhD-positive, so the anti-D channel is validated here for the positive call.