iBodies: a versatile tool for IVD diagnostics based on synthetic polymers Libor Kostka 1 , Vladimír Šubr 1 , Pavel Šácha 2 , Jan Konvalinka 2 , Tomáš Etrych 1 1 Institute of Macromolecular Chemistry CAS, Czechia, 2 Institute of Organic Chemistry and Biochemistry CAS, Czechia Monoclonal antibodies are widely used tools for research, diagnostics, and anticancer therapy. However, their use is compromised by high production costs, limited stability, and complicatedchemical modification. Recently,we reported the design and development of synthetic polymer conjugates capable of replacing antibodies in several in vitro applications used in biochemistry, such as ELISA, flow cytometry, immunocytochemistry, and immunoprecipitation, and also useful for specific imaging of tumors. The conjugates, named "iBodies", consist of a hydrophilic copolymer decorated with low-molecular-weight compounds that function as targeting ligands, affinity anchors (typically biotin), and imaging probes (fluorophores or radiolabels). 1,2 We chose and synthesized water-soluble and biocompatible N -(2-hydroxypropyl) metha-acrylamide (HPMA) copolymers since they have been used to develop drug delivery carriers, imaging agents, and polymer drugs. 3 Chosen carrier, the HPMA-based copolymer, is a multivalent synthetic macromolecule with many reactive groups which enable covalent attachment of various ligands. These polymers were synthesized by the controlled polymerization technique RAFT in the presence of a trithiocarbonate chain transfer agent in organic solvents. The molecular weights of prepared copolymers ranged from 35000 to 100000 g/mol with polydispersities below 1,2. The content of co-monomer units with reactive thiazolidine-2-thione groups was between 8 - 20 mol%. These reactive groups allow covalent attachment of different ligands bearing primary amino groups at common conditions and room temperature. We prepared specific conjugates targeting several important proteins, such as prostate-specific membrane antigen, fibroblast activating protein, and a number of other medicinally relevant enzymes. We used these iBodies for enzyme inhibition, protein isolation, immobilization, quantification, and live cell imaging. Our data indicate that this highly modular and versatile polymer system can produce inexpensive and stable antibody substitutes. This work was financially supported by the grant of Czech Science Foundation (grant No.23-05642S). References
1. Šácha P. et al. Angew Chem Int Ed, 2016, 55 (7), 2356-2360. 2. Dvořáková P. et al., J Med Chem, 2017, 60 (20), 8385-8393 3. Ulbrich K. et al., Chem Rev, 2016, 116 (9), 5338-5431
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