Efficient and Simple Paper-Based Assay for Plasma Separation Using Universal Anti-H Agglutinating Antibody

[Image: see text] Background: Conventional laboratory tests require plasma separation using centrifugation by skilled personnel in well-equipped lab. Development of a simple, reliable, and cheap point-of-care (POC) test for plasma separation will overcome these limitations. Methods: Plasma separation was achieved in filter paper using the anti-H agglutinating antibody. Hydrophobic channels were created using a solid ink printer. The reproducibility, efficiency, recovery, and applicability of the assay were validated on a large number of blood samples. Results: A simple, fast, cheap, and direct paper-based assay for plasma separation from whole blood using universal anti-H agglutinating antibody was developed without equipment or pretreatment requirements. The purity of plasma separation using anti-H treated paper was confirmed by microscopy and biuret test for plasma albumin detection. Plasma separation was affected by paper structure, antibody concentration, donor gender, and hematocrit. The efficiency of the assay was 72% and the reproducibility was about 90% with minimal interassay and intra-assay variabilities. The assay successfully separated plasma from 116/119 samples, indicating high sensitivity (97.5%). Furthermore, the assay accurately recovers thyroid stimulating hormone from samples compared to standard methods with 107% recovery rate. Conclusions: Paper-based plasma separation using anti-H agglutinating antibodies would have numerous applications in paper-based POC tests and in resource limited areas.