A bi-functional polymeric coating for the co-immobilization of proteins and peptides on microarray substrates


Posted: 2021-11-10 20:00:00
Anal Chim Acta . 2021 Dec 1;1187:339138. doi: 10.1016/j.aca.2021.339138. Epub 2021 Oct 9. Affiliations Expand Affiliations 1 Istituto di Scienze e Tecnologie Chimiche "G.Natta", National Research Council of Italy, Via Mario Bianco 9, 20131, Milan, Italy. Electronic address: laura.sola@scitec.cnr.it. 2 Istituto di Scienze e Tecnologie Chimiche "G.Natta", National Research Council of Italy, Via Mario Bianco 9, 20131, Milan, Italy. Item in Clipboard Laura Sola et al. Anal Chim Acta. 2021. Show details Display options Display options Format Anal Chim Acta . 2021 Dec 1;1187:339138. doi: 10.1016/j.aca.2021.339138. Epub 2021 Oct 9. Affiliations 1 Istituto di Scienze e Tecnologie Chimiche "G.Natta", National Research Council of Italy, Via Mario Bianco 9, 20131, Milan, Italy. Electronic address: laura.sola@scitec.cnr.it. 2 Istituto di Scienze e Tecnologie Chimiche "G.Natta", National Research Council of Italy, Via Mario Bianco 9, 20131, Milan, Italy. Item in Clipboard CiteDisplay options Display options Format Abstract The analytical performance of the microarray technique in screening the affinity and reactivity of molecules towards a specific target, is highly affected by the coupling chemistry adopted to bind probes to the surface. However, the surface functionality limits the biomolecules that can be attached to the surface to a single type of molecule, thus forcing the execution of separate analyses to compare the performance of different species in recognizing their targets. Here we introduce a new N, N-dimethylacrylamide-based polymeric coating, bearing simultaneously different functionalities (N-acryloyloxysuccinimide and azide groups) to allow an easy and straightforward method to co-immobilize proteins and oriented peptides on the same substrate. The bi-functional copolymer has been obtained by partial post polymerization modification of the functional groups of a common precursor. A NMR characterization of the copolymer was conducted to quantify the percentage of NAS that has been transformed into azido groups. The polymer was used to coat surfaces onto which both native antibodies and alkyne modified peptides were immobilized, to perform the phenotype characterization of extracellular vesicles (EVs). This strategy represents a convenient method to reduce the number of analysis, thus possible systematic or random errors, besides offering a drastic shortage in time, reagents and costs. Keywords: Click chemistry; Co-immobilization; Extracellular vesicles; Functional polymers; Peptide microarray; Protein microarray. Copyright © 2021 Elsevier B.V. All rights reserved. Conflict of interest statement Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. [x] Cite Copy Format: Send To [x]

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