title: Bioconjugation of peptides using advanced nanomaterials to examine their interactions in 3D printed flow-through device.
abstract: Peptide-peptide interactions are crucial in the living cell as they lead to the formation of the numerous types of complexes. In this study, synthetic peptides containing 11 of cysteines (α-domain of metallothionein (MT)) and sialic acid binding region (130-loop of hemagglutinin (HA)) were employed. The aim of the experiment was studying the interactions between MT and HA-derived peptides. For this purpose, fragments were tagged with cysteines at C-terminal part to serve as ligand sites for PbS and CuS quantum dots (QDs), and therefore these conjugates can be traced and quantified during wide spectrum of methods. As a platform for interaction, γ-Fe2O3 paramagnetic particles modified with tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane (hydrodynamic diameter 30-40 nm) were utilized and MT/HA interactions were examined using multi-instrumental approach including electrochemistry, electrophoretic methods, and MALDI-TOF/TOF mass spectrometry. It was found that peptides enter mutual creation of complexes, which are based on some of nonbonded interactions. The higher willingness to interact was observed in MT-derived peptides toward immobilized HA. Finally, we designed and manufactured flow-through electrochemical 3D printed device (reservoir volume 150 μL) and utilized it for automated analysis of the HA/MT metal labels. Under the optimal conditions, (deposition time and flow rate 80 s and 1.6 mL/min for CuS and 120 s and 1.6 mL/min PbS, respectively), the results of peptide-conjugated QDs were comparable with atomic absorption spectrometry.
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Michalek
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Lukas
authors.initials: L
authors.lastname: Richtera
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Krejcova
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Lukas
authors.initials: L
authors.lastname: Nejdl
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Renata
authors.initials: R
authors.lastname: Kensova
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Zitka
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Kopel
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Heger
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University, Brno, Czech Republic.|Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
authors.forename: Rene
authors.initials: R
authors.lastname: Kizek
mesh_terms: D058185:Magnetite Nanoparticles / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D066330:Printing, Three-Dimensional; D011485:Protein Binding
chemical_list: D058185:Magnetite Nanoparticles; D010455:Peptides
keywords: 3D printing; Advanced materials; Influenza; Lab-On-a-Chip; Magnetic particle