Microarray and Sensor Technology

The unit addresses biotechnological questions involving the interface between biological structures and technical systems. Special focus is aimed at gaining as much analytical information as possible from very small quantities of material. The solution lies in the modification of the respective surfaces.

Surfaces with new characteristics or a so-called intelligent surface can be produced by using defined (bio)chemical functionalization (branched linkers) or by applying thin films (biopolymer-based membranes or switchable hydrogel layers). The technology is implemented both on geometric materials such as fibers as well as on planar media such as plates or chips. The surfaces themselves vary from glass and wafer materials to plastics.

The products developed by the unit are independent sensor elements (test strip based PoNd) or analyses and database tools (cell and peptide chips) and can be employed in various applications in the areas of environmental analysis, food industry monitoring, herd management, process control or diagnostics.

Offerings of the Microarrays and Sensor Technology Unit

Functional Surface Structuring

  • Activation, modification and (polymer) synthesis of diverse surfaces
  • Targeted use of separation and permeation characteristics of thin layers
  • Optimization of the binding reactions by thermodynamic and kinetic measurement
  • Surface characterization (IR, MS, Ellipsometer)
  • Measurement of interaction characteristics (ITC, SPR, Nanotemper)

Real-time Monitoring

  • On-site monitoring of implementation, growth and/or chemical processes
  • Physicochemical characterisation of the interplay of biomolecules

Spatially-Resolved Analytics Based on DNA, RNA, Peptides, Proteins and Cells

  • Probe design
  • Physicochemical and biochemical analytics
  • Manufacture of customer-specific DNA, peptide and protein chips with diverse contact and non-contact spotters
  • Hundred percent monitoring of the spot process
  • Quality control with contact angle measurement and AFM
  • Execution of hybridising or other binding reactions with customer's material and evaluation of microarray experiments

Development of New High-Throughput Detection Tools

  • Specific enrichment with surface-modified micro- and nano-particles
  • Characterization of the permeation characteristics of thin layers

Multiparameter Analytics (including for target and active ingredient screening, host-pathogen interactions)

  • Transfer / miniaturization of existing analyses in alternative (sensor) systems
  • Pre-organized mimetica
  • Separation with structure- and surface-modified micro- and nano-particles
  • Development and characterization of bioanalogue receptors
  • Assay development

Minimum

As a part of a large-scale project for the rapid detection of pathogens, the work group has developed a high-definition molecular coating for new sensor materials to allow the differentiated and highly selective label-free detection of various pathogens.

The new sensors can be modified for use in coupling biological components such as antibodies, antigens, and peptides up to entire cells, and nevertheless not lose their optical characteristics.

However, the modifications are not permitted to entail any restrictions on the binding behaviour of the capture molecules, since this would adversely affect the subsequent analysis. The binding characteristics cannot be influenced through the loss of a degree of freedom.

Thanks: The project underlying this report was promoted with funds from the German Ministry for Economy and European Affairs of the State of Brandenburg and the EU.

Handkerchief Laboratory

The goal of the “handkerchief laboratory” is the development of a new generation of biosensors that specifically bind with the analyte and allow its detection without specific tools.

Special anchor structures produce multiple recognition domains that are adapted, combined and positioned so to efficiently bind to the pathogen to be recognised. This anchoring also serves as the functional connection to a substrate that additionally ensures signal coupling. Furthermore, the framework may not falsify the recognition reaction or endanger the inertness of the recognition domains vis-a-vis the coupling reaction. Depending on the selected signal coupling, both the chemical reaction as well as the sequence of additional steps (relative sequence of the assembly of recognition domains, signallers and substrates) are adapted and varied. Important problem-solving include the incorporation of suitable spacer groups for the recognition of domains and their relative positioning to each other, to signal-generating groups, and to the substrate.

Thanks: The project underlying this report was promoted with funds from the German Federal Ministry for Education and Research (03IS2201A).

TheraSense

The goal of the project is to develop autonomous biosensors and connect them to telecommunication units. The concept offers possibilities for integrating the necessary preanalytics with the sensor systems. It was based on exemplary medical scenarios to develop demonstrators for selected parameters.

Thanks: The project underlying this report was promoted with funds from the German Ministry for Economy and European Affairs of the State Brandenburg and the EU.

Biochip Manufacture

  • Contact and non-contact biochip arrayer for the production of DNA, peptides, proteins and additional biochips
  • Diverse biochip scanners: Tecan scanner, Applied Precision "Arrayworx"
  • In-house development “FLOW” for simultaneous kinetic measurements in flow-through

Biosensors

  • Label-free biosensors (BiacoreTM T100, BiacoreTM T200, Reichert SR7000DC, Monolith NT.115, MicroCal VP-ITC, Flow-chip calorimeter, ICT, MST, Ellipsometer)
  • Fiber optical immuno-assays sensor analyser
  • Flow cytometer
  • Electrochemical workstation (Amperometrics, CV, SWV, DPP, OCPT, etc.)
  • Nanoliter micro-dispenser
  • Climate chamber

Instrumental Analytics

  • HPLC
  • Mass spectrometry
  • FT-IR spectrometer
  • Fluorescence MTP reader
  • UV-NIR spectrophotometer
  • µL-UV-Vis spectrophotometer
  • UV-Vis spectroscopic MTP reader
  • Bio luminescence / fluorescence MTP reader and spectrometer

Publications

  • Bader D, Klier DT, Hettrich C, Bier FF, Wessig P. Detecting carbohydrate–lectin interactions using a fluorescent probe based on DBD dyes. Anal. Methods. 2016 Jan 13; 8, 1235-1238: DOI dx.doi.org/10.1039/C5AY02991K
  • Couturier J-P, Wischerhoff E, Benin R, Hettrich C, Koetz J, Sütterlin M, Tiersch B, Laschewsky A. Thermoresponsive Polymers and Inverse Opal Hydrogels for the Detection of Diols. Langmuir. 32 (2016) 4333-4345: DOI dx.doi.org/10.1021/acs.langmuir.6b00803
  • Prashant P, Elangovan E, Hettrich C, Möller HH, Linker T. Synthesis of 2-Thiocarbohydrates and Their Binding to Concanavalin A. J. Org. Chem. 81 (2016) 8595–8603: DOI dx.doi.org/10.1021/acs.joc.6b00987
  • Couturier J-P, Sütterlin M, Laschewsky A, Hettrich C, Wischerhoff E. Inverse Opale aus responsiven Hydrogelen für die Detektion von Makromolekülen. Angew. Chem. 127 (2015) 6741-6745, DOI dx.doi.org/10.1002/ange.201500674.
  • Couturier J-P, Sütterlin M, Laschewsky A, Hettrich C, Wischerhoff E. Responsive Inverse Opal Hydrogels for the Sensing of Macromolecules. Angew. Chem. Int. Ed. 54 (2015) 6641-6644, DOI dx.doi.org/10.1002/anie.201500674.
  • Hüttl C, Hettrich C, Riedel M, Henklein P, Rawel H, Bier FF. Development of peptidyl lysine dendrons: 1,3-Dipolar cycloaddition for peptide coupling and antibody recognition. Chemical Biology and Drug Design. 85 (2015) 565-573. DOI dx.doi.org/10.1111/cbdd.12444.
  • Kumke M, Eisold U, Sellrie F, Schenk J, Stöcklein W, Lenz C. Bright or dark immune complexes of anti-TAMRA antibodies for tailored fluorescence based bioanalysis. Anal. Bioanal. Chem. 2014 eingereicht.
  • Hovestädt M, Memczak H, Pleiner D, Zhang X, Rappich J, Bier FF, Stöcklein WFM. Characterization of a new maleimido functionalization of gold for surface plasmon resonance spectroscopy. J. Mol. Recognit. 2014, in press, DOI dx.doi.org/10.1002/jmr.2396.
  • Hüttl C, Hettrich C, Miller R, Paulke B-R, Henklein P, Rawel R, Bier FF. Self-assembled peptide amphiphiles function as multivalent binder with increased hemagglutinin affinity. BMC Biotechnology 2013, 13, 51 s.
  • Kozma P, Lehmann A, Wunderlich K, Michel D, Schumacher S, Ehrentreich-Förster E, Bier FF. A novel handheld fluorescent microarray reader for point-of-care diagnostic. Biosensors & bioelectronics 47 (2013), S.415-420.
  • Schumacher S, Lüdecke C, Ehrentreich-Förster E, Bier FF. Platform technologies for molecular diagnostics near the patient's bedside. Molecular diagnostics Berlin: Springer, 2013 (Advances in biochemical engineering/biotechnology 133) ISBN: 978-364-23769-1-7.
  • Dechtrirat D, Jetzschmann KJ, Stöcklein WFM, Scheller FW, Gajovic-Eichelmann N. Protein rebinding to a surfice-confined imprint. Adv. Funct. Mater. 22, 5231-7 (2012).
  • Stech M, Merk H, Schenk JA, Stöcklein W, Wüstenhagen D, Micheel B, Duschl C, Bier FF, Kubick S. Production of functional antibody fragments in a vesicle-based eukaryotic cell-free translation system. J. Biotechnol. (2012) 164, 220-231.
  • Heise C, Ehrentreich-Förster E, Bier FF. Dansyl, a fluorescent photoprotecting group for microarray applications. Journal of chemical technology and biotechnology : JCTB 87 (2012), Nr.11, S.1584-1592.
  • Grießner M, Hartig D, Christmann A, Pohl C, Schellhase M, Ehrentreich-Förster E. Development and characterization of a disposable plastic microarray printhead. Biomedical microdevices 13 (2011), Nr.3, S.533-538.
  • Schumacher S, Katterle M, Hettrich C, Paulke B-R, Hall DG, Scheller FW, Gajovic-Eichelmann N. Label-free Detection of Enhanced Saccharide binding at pH 7.4 to Nanoparticulate Benzoboroxole-based Receptor Units. Journal of Molecular Recognition 24 (2011) S. 953-959.
  • Tan C, Gajovic-Eichelmann N, Stöcklein WFM, Polzius R, Bier FF. Direct detection of Delta-9-Tetrahydrocannabinol in saliva using a novel homogeneous competitive immunoassay with fluorescence quenching. Anal. Chim. Acta 658, 187-192 (2010).
  • Nagel T, Ehrentreich-Förster E, Bier FF. Label-free serodiagnosis on a grating coupler. Rasooly, A.; Herold, K.: Biosensors and Biodetection: Methods and Protocols. Volume 1: Optical-Based Detectors Totowa, N.J.: Humana Press, 2009 (Methods in molecular biology 2256) ISBN: 978-1-60327-566-8 Kap.10.
  • Ehrentreich-Förster E, Orgel D, Krause-Griep A, Cech B, Erdmann VA, Bier F, Scheller FW, Rimmele M. Biosensor-based on-site explosives detection using aptamers as recognition elements. Analytical and bioanalytical chemistry 391 (2008), Nr.5, S.1793-1800.
  • Nagel T, Ehrentreich-Förster E, Singh M, Schmitt K, Brandenburg A, Berka A, Bier FF. Direct detection of tuberculosis infection in blood serum using three optical label-free approaches. Sensors and Actuators B: Chemical, Volume 129, Issue 2, 2008, 934-940.
  • Antwerpen MH, Schellhase M, Ehrentreich-Förster E, Bier FF, Witte W, Nübel U. DNA microarray for detection of antibiotic resistance determinants in Bacillus anthracis and closely related Bacillus cereus. Molecular and cellular probes, Vol.21 (2007), No.2, pp.152-160.
  • Andresen H, Grötzinger C, Zarse K, Birringer M, Hessenius C, Kreuzer OJ, Ehrentreich-Förster E, Bier FF. Peptide microarrays with site-specifically immobilized synthetic peptides for antibody diagnostics. Sens Actuators B Chem 2005 Volume 113, Issue 2, 27 February 2006, Pages 655-663.
  • Andresen H, Zarse K, Grötzinger C, Kreuzer OJ, Ehrentreich-Förster E, Bier FF. Functional Peptide Microarrays for Specific and Sensitive Antibody Diagnostics. Proteomics 6 (2006), Nr.5, S.1376-1384.

Patents

  • Reiss E, Bier F, Stöcklein W. Signalgebende Bindemoleküle, Vorrichtung und Verfahren zu deren Verwendung. DE 10 2010 010 052 A1. 8. Nov. 2011.
  • Bier F, Ehrentreich-Förster E. Koimmibilisierung mehrerer chemischer Spezies. DE 100 02 895 B4. 9. Feb. 2006.
  • Hettrich C, Hettrich K, Ehrentreich-Förster E. Herstellung von transparenten Filmen aus Cellulose-Dispersionen und deren Verwendung als multifunktionelle Träger für Liganden (Analyten).
  • Ehrentreich-Förster E, Kozma P, Schumacher S. Messgerät zur Lumineszenzmessung. DE 10 2012 018 303 A1. 20. März 2014.