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Highly Efficient Synthesis and Assay of Protein-Imprinted Nanogels by Using Magnetic Templates.


ABSTRACT: We report an approach integrating the synthesis of protein-imprinted nanogels ("plastic antibodies") with a highly sensitive assay employing templates attached to magnetic carriers. The enzymes trypsin and pepsin were immobilized on amino-functionalized solgel-coated magnetic nanoparticles (magNPs). Lightly crosslinked fluorescently doped polyacrylamide nanogels were subsequently produced by high-dilution polymerization of monomers in the presence of the magNPs. The nanogels were characterised by a novel competitive fluorescence assay employing identical protein-conjugated nanoparticles as ligands to reversibly immobilize the corresponding nanogels. Both nanogels exhibited Kd <10?pM for their respective target protein and low cross-reactivity with five reference proteins. This agrees with affinities reported for solid-phase-synthesized nanogels prepared using low-surface-area glass-bead supports. This approach simplifies the development and production of plastic antibodies and offers direct access to a practical bioassay.

SUBMITTER: Mahajan R 

PROVIDER: S-EPMC6471088 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Highly Efficient Synthesis and Assay of Protein-Imprinted Nanogels by Using Magnetic Templates.

Mahajan Rashmi R   Rouhi Mona M   Shinde Sudhirkumar S   Bedwell Thomas T   Incel Anil A   Mavliutova Liliia L   Piletsky Sergey S   Nicholls Ian A IA   Sellergren Börje B  

Angewandte Chemie (International ed. in English) 20181115 3


We report an approach integrating the synthesis of protein-imprinted nanogels ("plastic antibodies") with a highly sensitive assay employing templates attached to magnetic carriers. The enzymes trypsin and pepsin were immobilized on amino-functionalized solgel-coated magnetic nanoparticles (magNPs). Lightly crosslinked fluorescently doped polyacrylamide nanogels were subsequently produced by high-dilution polymerization of monomers in the presence of the magNPs. The nanogels were characterised b  ...[more]

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