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Nanobiotechnology - Petr Skládal

Research Group Leader
Researcher ID
Phone: +420 54949 7659, +420 54949 7010
E-mail: ,
Office:
Deputy Group Leader
Phone: +420 54949 5606
E-mail: ,
Office:

Research areas

  • Imaging of biomolecules, cells and other biological objects using scanning probe microscopies (SPM – including AFM, SNOM and STM)
  • Characterization of affinity interactions using biosensors in real time
  • Development of biosensors using electrochemical, optical and piezoelectric transducers
  • Immobilization, modification and conjugation of biomolecules

Main objectives

  • Visualization and modification of biological objects including tissues, cells, cellular structures, and single biomolecules
  • Development of new methodologies for investigating the structure, interactions, and dynamics of biomolecules.
  • Investigations into the structure and interactions of biomacromolecules and their relation to the functions of living systems, disease and therapy.
  • Investigations into the behaviour of natural and chemically modified biomacromolecules at electrically charged surfaces linked to the development of novel electrochemical biosensors and bioassays.

Content of research

Nanobiotechnology represents advanced scanning probe microscopic techniques, nano-lithographic machining and various types of artificial nanostructures applied for either visualization or modification of biological objects including tissues, cells, cellular structures and biomolecules. The unique opportunity to touch a single individual molecule of a protein or nucleic acid with the scanning tip provides high-resolution sub-nanometer and pseudo-3-dimensional images providing details of such bioobjects in their native state. These approaches are currently revolutionizing many fields of biology, biophysics and biochemistry and provide innovative results and methodologies for applications in health care – nanobiosensing systems, nanoparticles for labelling and smart distribution of drugs (nanomedicine). Within this work package, the following research fields will be addressed:

Imaging of biosurfaces and biomolecules using scanning probe microscopies

The laboratory performs atomic force microscopy (AFM) scans in both dry state and in liquids in contact and non-contact modes. Modified tips allow characterization of surface hydrophobicity, and specific target molecules and cellular surfaces (e.g. tips modified with antibodies) are scanned (spectroscopic techniques) while conductive tips with applied potential will be used for bioelectrochemical studies. Repeated scans will allow the movement and morphologic changes of cells to be studied (VideoAFM). Supplementary information about cells and cellular elements will be provided by scanning near optical field microscopy (SNOM, overcoming the diffraction limit); and scanning tunneling microscopy (STM) will be chosen for atomic resolution.

skladal

Molecule of immunoglobulin G imaged on the mica surface using the cantilever with a diamond-like carbon tip, semicontact mode, Ntegra Vita atomic force microscope.

 

Nanobiointeractions and the measurement of forces within biocomplexes

The binding of two individual complementary molecules (antibody-antigen, ligand-receptor, and hybridization of oligonucleotides) will be studied using one partner bound to the solid support and the other linked to the scanning tip. The force data measured at the nanolevel will be correlated with the results obtained at the macrolevel using surface plasmon resonance techniques providing information about kinetics of affinity interactions in real time.

Nanomanipulation and nanolithography of biological objects

In the AFM-lithography mode, the tip of the cantilever in contact with the scanned object can be used to manipulate cells on the surface. Nano(bio)sensing arrays and other functional nanoobjects will be constructed.

Nanobiosensors and biosensing nanoarrays

Nanobiosensors will employ the cantilever as nanomechanical transducer bending due to the affinity interaction on one of its sides. Nanoarrays – biochips consisting of sets of specific recognition proteins (monoclonal or recombinant antibodies, engineered receptors and enzymes, artificial peptide folds designed by molecular modelling) will be incubated with clinical samples and then the binding pattern will be read with the help of AFM (SNOM, STM), either directly or after suitable amplification (magnetic nanoparticles, quantum dots). For the validation, “larger” micrometer-sized array elements will be produced and evaluated using multichannel SPR, fluorescence scanning and scanning electrochemical microscopy (SECM).

list / cards

Name and position

E-mail

Phone

Petr Skládal
Research Group Leader
+420 54949 7659, +420 54949 7010
Karel Lacina, Ph.D.
Researcher
+420 54949 7611
Jan Přibyl, Ph.D.
Researcher
+420 54949 5606
Veronika Horáčková
PhD student
+420 54949 7676
Matěj Pastucha
PhD student
+420 54949 8517
Zdeněk Farka
PhD student
+420 54949 7674
Ondřej Kubesa
PhD student
Zuzana Svozilová
PhD student
Veronika Čunderlová
odborná pracovnice

SELECTED PUBLICATIONS

2017

  • FARKA, Z; JURIIK, T; KOVAAR, D; TRNKOVA, L; SKLADAL, P, 2017:Nanoparticle-Based Immunochemical Biosensors and Assays: Recent Advances and Challenges. CHEMICAL REVIEWS
  • FARKA, Z; MICKERT, MJ; HLAVACEK, A; SKLADAL, P; GORRIS, HH, 2017:Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers. ANALYTICAL CHEMISTRY 89 (21), p. 11825 - 11830.
  • HLAVACEK, A; PETEREK, M; FARKA, Z; MICKERT, MJ; PRECHTL, L; KNOPP, D; GORRIS, HH, 2017:Rapid single-step upconversion-linked immunosorbent assay for diclofenac. MICROCHIMICA ACTA 184 (10), p. 4159 - 4165.
  • LACINA, K; KUBESA, O; VANYSEK, P; HORACKOVA, V; MORAVEC, Z; SKLADAL, P, 2017:Selective electrocatalysis of reduced graphene oxide towards hydrogen peroxide aiming oxidases-based biosensing: Caution while interpreting. ELECTROCHIMICA ACTA 223 , p. 1 - 7.
  • NARDONE, G; LA CRUZ, JOD; VRBSKY, J; MARTINI, C; PRIBYL, J; SKLADAL, P; PESL, M; CALUORI, G; PAGLIARI, S; MARTINO, F; MACECKOVA, Z; HAJDUCH, M; SANZ-GARCIA, A; PUGNO, NM; STOKIN, GB; FORTE, G, 2017:YAP regulates cell mechanics by controlling focal adhesion assembly. NATURE COMMUNICATIONS 8
  • SKARKOVA, P; NOVOTNY, K; LUBAL, P; JEBAVA, A; PORIZKA, P; KLUS, J; FARKA, Z; HRDLICKA, A; KAISER, J, 2017:2d distribution mapping of quantum dots injected onto filtration paper by laser-induced breakdown spectroscopy. SPECTROCHIMICA ACTA PART B-ATOMIC SPECTROSCOPY 131 , p. 107 - 114.

2016

  • CUNDERLOVA, V; HLAVACEK, A; HORNAKOVA, V; PETEREK, M; NEMECEK, D; HAMPL, A; EYER, L; SKLADAL, P, 2016:Catalytic nanocrystalline coordination polymers as an efficient peroxidase mimic for labeling and optical immunoassays. MICROCHIMICA ACTA 183 (2), p. 651 - 658.
  • FARKA, Z; JURIK, T; PASTUCHA, M; KOVAR, D; LACINA, K; SKLADAL, P, 2016:Rapid Immunosensing of Salmonella Typhimurium Using Electrochemical Impedance Spectroscopy: the Effect of Sample Treatment. ELECTROANALYSIS 28 (8), p. 1803 - 1809.
  • FARKA, Z; JURIK, T; PASTUCHA, M; SKLADAL, P, 2016:Enzymatic Precipitation Enhanced Surface Plasmon Resonance Immunosensor for the Detection of Salmonella in Powdered Milk. ANALYTICAL CHEMISTRY 88 (23), p. 11830 - 11836.
  • HLAVACEK, A; FARKA, Z; HUBNER, M; HORNAKOVA, V; NEMECEK, D; NIESSNER, R; SKLADAL, P; KNOPP, D; GORRIS, HH, 2016:Competitive Upconversion-Linked Immunosorbent Assay for the Sensitive Detection of Diclofenac. ANALYTICAL CHEMISTRY 88 (11), p. 6011 - 6017.
  • JURIK, T; PODESVA, P; FARKA, Z; KOVAR, D; SKLADAL, P; FORET, F, 2016:Nanostructured gold deposited in gelatin template applied for electrochemical assay of glucose in serum. ELECTROCHIMICA ACTA 188 , p. 277 - 285.
  • LACINA, K; ZAK, J; SOPOUSEK, J; SZABO, Z; VACLAVEK, T; ZERAVIK, J; FIALA, P; SKLADAL, P, 2016:Transistor Amplifier as an Electrochemical Transducer with Intuitive Optical Read-out: Improving Its Performance with Simple Electronic Solutions. ELECTROCHIMICA ACTA 216 , p. 147 - 151.
  • MAKHNEVA, E; MANAKHOV, A; SKLADAL, P; ZAJICKOVA, L, 2016:Development of effective QCM biosensors by cyclopropylamine plasma polymerization and antibody immobilization using cross-linking reactions. SURFACE & COATINGS TECHNOLOGY 290 , p. 116 - 123.
  • MANAKHOV, A; MAKHNEVA, E; SKLADAL, P; NECAS, D; CECHAL, J; KALINA, L; ELIAS, M; ZAJICKOVA, L, 2016:The robust bio-immobilization based on pulsed plasma polymerization of cyclopropylamine and glutaraldehyde coupling chemistry. APPLIED SURFACE SCIENCE 360 , p. 28 - 36.
  • PESL, M; PRIBYL, J; ACIMOVIC, I; VILOTIC, A; JELINKOVA, S; SALYKIN, A; LACAMPAGNE, A; DVORAK, P; MELI, AC; SKLADAL, P; ROTREKL, V, 2016:Atomic force microscopy combined with human pluripotent stem cell derived cardiomyocytes for biomechanical sensing. BIOSENSORS & BIOELECTRONICS 85 , p. 751 - 757.
  • SEDLMEIER, A; HLAVACEK, A; BIRNER, L; MICKERT, MJ; MUHR, V; HIRSCH, T; CORSTJENS, PLAM; TANKE, HJ; SOUKKA, T; GORRIS, HH, 2016:Highly Sensitive Laser Scanning of Photon-Upconverting Nanoparticles on a Macroscopic Scale. ANALYTICAL CHEMISTRY 88 (3), p. 1835 - 1841.
  • ZERAVIK, J; FOHLEROVA, Z; MILOVANOVIC, M; KUBESA, O; ZEISBERGEROVA, M; LACINA, K; PETROVIC, A; GLATZ, Z; SKLADAL, P, 2016:Various instrumental approaches for determination of organic acids in wines. FOOD CHEMISTRY 194 , p. 432 - 440.

2015

  • FARKA, Z; KOVAR, D; SKLADAL, P, 2015:Rapid Detection of Microorganisms Based on Active and Passive Modes of QCM. SENSORS 15 (1), p. 79 - 92.
  • LACINA, K; SKLADAL, P, 2015:Bipolar transistor amplifier for transduction of electrochemical response to visual perception. SENSORS AND ACTUATORS B-CHEMICAL 210 , p. 183 - 189.
  • SUN, XL; LACINA, K; RAMSAMY, EC; FLOWER, SE; FOSSEY, JS; QIAN, XH; ANSLYN, EV; BULL, SD; JAMES, TD, 2015:Reaction-based Indicator displacement Assay (RIA) for the selective colorimetric and fluorometric detection of peroxynitrite. CHEMICAL SCIENCE 6 (5), p. 2963 - 2967.

2014

  • HLAVACEK, A; SEDLMEIER, A; SKLADAL, P; GORRIS, HH, 2014:Electrophoretic Characterization and Purification of Silica-Coated Photon-Upconverting Nanoparticles and Their Bioconjugates. ACS APPLIED MATERIALS & INTERFACES 6 (9), p. 6930 - 6935.
  • KOVAR, D; FARKA, Z; SKLADAL, P, 2014:Detection of Aerosolized Biological Agents Using the Piezoelectric Immunosensor. ANALYTICAL CHEMISTRY 86 (17), p. 8680 - 8686.
  • MANAKHOV, A; SKLADAL, P; NECAS, D; CECHAL, J; POLCAK, J; ELIAS, M; ZAJICKOVA, L, 2014:Cyclopropylamine plasma polymers deposited onto quartz crystal microbalance for biosensing application. PHYSICA STATUS SOLIDI A-APPLICATIONS AND MATERIALS SCIENCE 211 (12), p. 2801 - 2808.
  • PESL, M; IVANA, A; PRIBYL, J; HEZOVA, R; VILOTIC, A; AIMOND, F; FAUCONNIER, J; VRBSKY, J; KRUZLIAK, P; SKLADAL, P; KARA, T; ROTREKL, V; LACAMPAGNE, A; DVORAK, P; MELI, AC, 2014:Molecular and Functional Characterization of Uniform-Sized Beating Embryoid Bodies and Cardiomyocytes from Human Embryonic and Induced Pluripotent Stem Cells. BIOPHYSICAL JOURNAL 106 (2), p. 565A - 565A.

2013

  • FARKA, Z; KOVAR, D; SKLADAL, P, 2013:PIEZOELECTRIC BIOSENSOR COUPLED TO CYCLONE AIR SAMPLER FOR DETECTION OF MICROORGANISMS. CHEMICKE LISTY 107 , p. S302 - S304.

2012

  • HLAVACEK, A; SKLADAL, P, 2012:Isotachophoretic purification of nanoparticles: Tuning optical properties of quantum dots. ELECTROPHORESIS 33 (42286), p. 1427 - 1430.
  • SKLADAL, P; SVABENSKA, E; ZERAVIK, J; PRIBYL, J; SISKOVA, P; TJARNHAGE, T; GUSTAFSON, I, 2012:Electrochemical Immunosensor Coupled to Cyclone Air Sampler for Detection of Escherichia coli DH5 alpha in Bioaerosols. ELECTROANALYSIS 24 (3), p. 539 - 546.

2011

  • NAGY, L; ANGYAL, T; NAGY, G; AKIKO, M; PRIBYL, J; SKLADAL, P, 2011:SENSITIVE ELECTROCHEMICAL METHOD DEVELOPMENT For "in vivo" Measurement of ROS in Ethanol Induced Stress. BIODEVICES 2011 , p. 359 - 362.

2010

  • CRHA, I; PRIBYL, J; SKLADAL, P; ZAKOVA, J; VENTRUBA, P; POHANKA, M, 2010:Determination of the surface pathology of human sperm by atomic force microscopy. HUMAN REPRODUCTION 25 , p. I122 - I122.
  • ZERAVIK, J; LACINA, K; JILEK, M; VLCEK, J; SKLADAL, P, 2010:Biosensor for determination of carboxylic acids in wines based on the inhibition of sarcosine oxidase. MICROCHIMICA ACTA 170 (3 IV), p. 251 - 256.

GRANTY

  • BIOMIMIC – Biomimetic sensors as a new generation of biotechnological devices for food safety and quality monitoring (230849), FP7- People - Marie Curie actions - IRSES, 2009 - 2012
  • Immunoassay for rapid diagnostics of European foulbrood (TJ01000386), TAČR ZETA, 2017 - 2019
  • Interactions of mutant forms of cryptogein with a membrane binding site: biosensors-based characterization (GAP501/11/1003), Czech Science Foundation - Standard Grants, 2011 - 2015
  • Nanobiotechnologies and biosensor for biointeraction studies – openning up the modern technology to researchers in biology (CZ.1.07/2.3.00/09.0167), MEYS - OP Education for Competiteveness, 2009 - 2012

CURRENT RESEARCH INFRASTRUCTURE

The infrastructure currently available includes Ntegra Vita AFM, Ntegra Solaris SNOM, Biacore 2000 SPR system, fluorescence microscope, cell cultivation facility, electrochemical (10), piezoelectric (5) and fiber optic (2) detectors, light sources (4), DAD spectrophotometer, Multiskan RC and Synergy 2 plate readers, autoinjectors (2), FIALab 3000, and data acquisition systems (NI). The group members have programming experience in Delphi, C++, Java and LabView.

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