A bi-functional polymeric coating for the co-immobilization of proteins and peptides on microarray substrates
| Autor: | Marina Cretich, Alessandro Mussida, Dario Brambilla, Roberto Consonni, Francesco Damin, Alessandro Gori, Marcella Chiari, Laura Sola |
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| Rok vydání: | 2021 |
| Předmět: |
Azides
Polymers Surface Properties 02 engineering and technology engineering.material 010402 general chemistry Biochemistry 01 natural sciences Analytical Chemistry Polymerization 03 medical and health sciences chemistry.chemical_compound Coating Environmental Chemistry Spectroscopy 030304 developmental biology chemistry.chemical_classification 0303 health sciences peptide microarray extracellular vesicles Biomolecule Click chemistry functional polymers co-immobilization protein microarray Polymer 021001 nanoscience & nanotechnology Microarray Analysis Combinatorial chemistry 0104 chemical sciences chemistry Alkynes engineering Protein microarray Peptide microarray Azide Functional polymers 0210 nano-technology Peptides |
| Zdroj: | ChemRxiv Analytica chimica acta 1187 (2021). doi:10.1016/j.aca.2021.339138 info:cnr-pdr/source/autori:Sola L.; Brambilla D.; Mussida A.; Consonni R.; Damin F.; Cretich M.; Gori A.; Chiari M./titolo:A bi-functional polymeric coating for the co-immobilization of proteins and peptides on microarray substrates/doi:10.1016%2Fj.aca.2021.339138/rivista:Analytica chimica acta (Print)/anno:2021/pagina_da:/pagina_a:/intervallo_pagine:/volume:1187 |
| ISSN: | 1873-4324 |
| Popis: | The analytical performance of the microarray technique in screening the affinity and reactivity of several probes 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 (DNA, protein, or peptide), 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 (NAS) of a common precursor. A deep characterization of the copolymer was carried out by means of NMR to quantify the percentage of NAS that has been transformed into azido groups. The polymer was then used to coat surfaces onto which both native antibodies and alkyne modified peptides were immobilized, to perform the phenotype characterization of extracellular vesicles (EVs). Ultimately, 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. |
| Databáze: | OpenAIRE |
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