Analytical Devices Based on Direct Synthesis of DNA on Paper

Abstract

This paper addresses a growing need in clinical diagnostics for parallel, multiplex analysis of biomarkers from small biological samples. It describes a new procedure for assembling microarrays of ssDNA and proteins on paper. This method starts with the synthesis of DNA oligonucleotides covalently linked to paper, and proceeds to generate DNA arrays capable of simultaneously capturing DNA, DNA-conjugated protein antigens, and DNA-conjugated antibodies. The synthesis of ssDNA oligonucleotides on paper is convenient and effective, with32% of the oligonucleotides cleaved and eluted from the paper substrate being full-length by HPLC for a 32-mer. These ssDNA arrays can be used to detect fluorophore-linked DNA oligonucleotides in solution, and as the basis for DNA-directed assembly of microarrays of DNA-conjugated capture antibodies on paper, detect protein antigens by sandwich ELISAs. Paper-anchored ssDNA arrays with different sequences can be used to assemble paper-based devices capable of detecting DNA and antibodies in the same device, and enable simple microfluidic paperbased devices.3

Autocatalytic Metallization of Fabrics Using Si Ink, for Biosensors, Batteries and Energy Harvesting

“…We have fabricated low‐cost electrochemical microfluidic paper‐based analytical devices (µPADs) with dual porous Ag working electrode (WE) and counter electrode (CE) formed with the SIAM process, as shown in Figure A . The Ag/AgCl pseudo‐reference electrode (RE) was printed on the rear side of a separate sheet of paper using a commercial ink and laminated immediately under the WEs (Figure B).…”

Orientation and characterization of immobilized antibodies for improved immunoassays (Review)

“…The DDI immobilized antibody was found to be an order of magnitude more sensitive to the EBOV GP antigen. Glavan et al 90 synthesized single-stranded DNA onto paper substrates and investigated antihuman C-reactive protein (hCRP) immobilized via DDI for binding hCRP from serum in a sandwich ELISA. However, DNA conjugation to the antibody utilized nonsite-directed NHS chemistry.…”

Coated and uncoated cellophane as materials for microplates and open-channel microfluidics devices

“…The unmodified surface properties of uncoated cellophane, which is chemically similar to paper, may also provide a platform for cell-culture, or for analytical applications requiring immobilization of DNA or proteins on its surface. 30,31 We used polyurethane molds with patterns analogous to those of a 96-well plate to emboss wells into uncoated cellophane C OH (see Figure S3). Wells embossed in dry cellophane C OH buckled and deformed when we placed them in contact with water, since water swells uncoated cellophane.…”