In this work, the preparation of glucose encapsulating nanoliposomes was achieved using two different lipid formulations, labelled as F1 and F2. Both formulations contained phosphatidylcholine (PC), oleylamido-4-butanoic acid (OABA) and in addition, F1 had cholesterol (CHO) while F2 contained cholesteroyl hemisussinate (CHEMS). These formulations were studied for their pH sensitivity and controlled release of encapsulated glucose for indirect detection of prostate-specific antigen (PSA) using sandwich immunoassay. As a signal generator, encapsulated glucose in nanoliposomes was quantified directly using the personal glucose meter (PGM) and colorimetrically using peroxidase property of horseradish peroxidase (HRP) enzyme and Pd|PdO as nanozymes. Controlled release of the encapsulated glucose was achieved using the pH effect or Triton-X 100 as a surfactant to destabilize the liposomal structure. The F2 formulation showed maximum controlled release at acidic phosphate buffer saline (PBS, pH 5.0). The concentration of encapsulated glucose was found to be high in F2 formulation and these were applied for the indirect detection of PSA. The limit of detection (LOD) values for PSA were found to be 53 fg mL-1, 64 fg mL-1 and 10 fg mL-1 when HRP, Pd|PdO and PGM were respectively used. The detection signal was linear over a wide concentration range for PSA including the clinical range of 4-10 ng mL-1. The HRP system showed low LOD value when compared with Pd|PdO nanozymes. PGM system gave lowest LOD values owing to the sensitivity of the system towards glucose. Pd|PdO nanozyme system showed good stability over a wide temperature up to 80 °C. PGM system required less reaction time (2 min), low reagents and results were readily generated in digital format.
Keywords: Controlled release; Glucose detection; Indirect immunoassay; Nanoliposomes; Nanozymes; Prostate-specific antigen.
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