Efficient in situ growth of enzyme-inorganic hybrids on paper strips for the visual detection of glucose

- Eliminated the need for centrifugation and dry process as the solution was drop-casted onto the paper.
- The accuracy and stability of pipetting resulted in a highly reproducible sensing material, which contributes to the reliability of the μPAD.
- The intimate contact of the natural and artificial enzymes enhanced the efficiency and sensitivity for clinical sample analysis.

A visual colorimetric microfluidic paper-based analytical device (μPAD) was constructed following the direct synthesis of enzyme-inorganic hybrid nanomaterials on the paper matrix. An inorganic solution of MnSO4 and KH2PO4 containing a diluted enzyme (glucose oxidase, GOx) was subsequently pipetted onto cellulose paper for the in situ growth of GOx@Mn3(PO4)2 hybrid functional materials. The characterization of the morphology and chemical composition validated the presence of hybrid materials roots in the paper fiber, while the Mn3(PO4)2 of the hybrid provided both a surface for enzyme anchoring and a higher peroxidase-like catalytic activity as compared to the Mn3(PO4)2 crystal that was synthesized without enzyme modulation. This new approach for the in situ growth of an enzyme-inorganic hybrid on a paper matrix eliminates centrifugation and the dry process by casting the solution on paper. The sensing material loading was highly reproducible because of the accuracy and stability of pipetting, which eventually contributed to the reliability of the μPAD. The self-assembled natural and artificial enzyme hybrid on the μPADs specifically detected glucose from a group of interferences, which shows great specificity using this method. Moreover, the colorimetric signal exhibited detection limitation for glucose is 0.01 mM, which lies in the physiological range of glucose in biological samples.