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Highly Efficient Synthesis and Assay of Protein‐Imprinted Nanogels by Using Magnetic Templates
Author(s) -
Mahajan Rashmi,
Rouhi Mona,
Shinde Sudhirkumar,
Bedwell Thomas,
Incel Anil,
Mavliutova Liliia,
Piletsky Sergey,
Nicholls Ian A.,
Sellergren Börje
Publication year - 2019
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201805772
Subject(s) - template , chemistry , polymerization , magnetic nanoparticles , nanoparticle , fluorescence , combinatorial chemistry , conjugated system , nanogel , monomer , solid phase synthesis , chromatography , polymer , nanotechnology , materials science , drug delivery , peptide , organic chemistry , biochemistry , physics , quantum mechanics
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 K d <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.