Inspired by biosystems, a process is proposed for preparing next-generation artificial polymer receptors with molecular recognition abilities capable of programmable site-directed modification following construction of nanocavities to provide multi-functionality. The proposed strategy involves strictly regulated multi-step chemical modifications: 1) fabrication of scaffolds by molecular imprinting for use as molecular recognition fields possessing reactive sites for further modifications at pre-determined positions, and 2) conjugation of appropriate functional groups with the reactive sites by post-imprinting modifications to develop programmed functionalizations designed prior to polymerization, allowing independent introduction of multiple functional groups. The proposed strategy holds promise as a reliable, affordable, and versatile approach, facilitating the emergence of polymer-based artificial antibodies bearing desirable functions that are beyond those of natural antibodies.

Novel sequential post-imprinting modifications were demonstrated on the development of multi-functionalized molecularly imprinted polymers for a biomarker glycoprotein.

Inspired by biosystems,ap rocess is proposed for preparing next-generation artificial polymer receptors with molecular recognition abilities capable of programmable sitedirected modification following construction of nanocavities to providem ulti-functionality.T he proposed strategy involves strictly regulated multi-step chemical modifications:1 )fabrication of scaffolds by molecular imprinting for use as molecular recognition fields possessing reactive sites for further modifications at pre-determined positions,and 2) conjugation of appropriate functional groups with the reactive sites by post-imprinting modifications to develop programmed functionalizations designed prior to polymerization, allowing independent introduction of multiple functional groups.T he proposed strategy holds promise as areliable,affordable,a nd versatile approach,f acilitating the emergence of polymerbased artificial antibodies bearing desirable functions that are beyond those of natural antibodies.

We report simultaneous detection of tumor marker proteins using a molecularly imprinted polymer-based fluorescence sensing system, in which prostate-specific antigen (PSA) recognition cavity, labeled with Alexa Fluor 594, and α-fetoprotein (AFP) recognition cavity, labeled with Alexa Fluor 647, exist together in the polymer matrix. The individually fluorescent-labeled PSA- and AFP-imprinted polymer was prepared by a dual imprinting method, followed by multi-step post-imprinting modifications (PIM). A polymerizable group, conjugated with PSA or AFP via a disulfide bond, was prepared and immobilized on a phenylboronic acid moiety-introduced substrate by the formation of cyclic diester between phenylboronic acid and glycans on proteins. The polymer matrix was prepared using surface-initiated atom transfer radical polymerization. After the reduction of the disulfide bond and hydrolysis of the cyclic diester, PSA- and AFP-imprinted nano-cavities were generated simultaneously. In multi-step PIM, thiol-reactive fluorescent dyes were introduced via a dynamic protection procedure using the target protein, which yielded dual fluorescence-labeled imprinted nano-cavities. Fluorescence signaling abilities were assessed, and each AFP and PSA-imprinted nano-cavity was confirmed to transduce the protein binding events into specific fluorescence signals, with lower values of limit of detection (<2.0 ng/mL). Therefore, the proposed methodology could be a novel platform for the simultaneous detection of multiple proteins.