Immunoanalytics

The Division at a glance

In the Division Immunoanalytics everything spins around antibodies and their analytical use. These Y-shaped molecules introduce a high selectivity in our methods which encompass any type of antibody-based analytical method.

Besides the application of antibodies we also work on improvements while raising monoclonal antibodies.

Another key activity is the development of validation concepts and issues of quality assurance with immunoassays, e.g. calibration and sound determination of the limit of detection. We conduct comparative studies with instrumental reference methods, which we develop ourselves. The developed and validated immunoreagents and conjugates are provided via the BAM webshop.

The staff members consider themselves as contact persons for researchers, developers and users in the sense of a competence centre immunoassay.

Key activities

Liquid handling workstation for pipetting in a microtiter plate

• Development of monoclonal antibodies (in cooperation with companies)
• Hapten design, synthesis und characterisation by NMR and x-ray structural analysis (in cooperation with Division 1.3)
• Conjugate synthesis (carrier proteins, enzymes) and conjugate characterisation by ESI-MS, MALDI-TOF-MS and CE
• Establishment and optimisation of immunoassays of various formats: enzyme immunoassay (ELISA), fluorescence immunoassay (FIA), fluorescence polarisation immunoassay (FPIA), chemiluminescence immunoassay (CLEIA)
• Development of validation concepts for immunoassays e.g. in-house validation, uncertainty budgets, best calibration practice, limit of detection
• Development of materials for immunoaffinity extraction
• Referece analysis: LC-MS/MS, HR-MS (Orbitrap), eventually after solid-phase extraction (SPE)
• Delivery of standardised immunoreagents (antibodies and tracer/conjugates)
• Analysis of samples for customers by ELISA or LC-MS/MS
• Member of the Bioanalysis Working Group of CCQM; participation in interlaboratory comparison studies

Equipment and methods

Photometer for microtiter plates

• Orbitrap-HRMS workstation “Exactive” with Capillary LC (Thermo Scientific) - shared with Divisions 1.5, 1.9, 1.10
• LC-MS/MS workstation with TSQ (triple stage quadrupole), allowing ESI/APCI ionisation (API 4000, AB Sciex)
• HPLC-UV/vis/FLU workstation (System 1200, Agilent) with fraction collector
• MALDI-TOF/TOF-MS (Bruker Autoflex III) - shared with 1.0
• MALDI-TOF-MS (Bruker Reflex III) - shared with 1.0
• Solid phase extraction workstation AutoTrace (Caliper)
• Capillary electrophoresis (CE) workstation (Agilent)
• Multidetection microplate reader (SpectraMax M5, Molecular Devices) - shared with Division 1.10
• Microtiterplate absorbance reader (SpectraMax Plus384, Molecular Devices)
• Microtiterplate 96-channel washer (ELx405™, BioTek)
• Microtiterplate 96-channel pipeting station (Liquidator96, Rainin)
• Microtiterplate liquid handling workstation epMotion (Eppendorf)
• Isothermal titration calorimeter VP-ITC (MicroCal) - shared with Division 1.5
• ASE Accelerated Solvent Extraction (Dionex)
• Cell culture laboratory with 2 laminar flow benches and incubator
• Flow cytometer CyFlow space (Partec) with particle and cell sorting unit, two laser sources (488 nm and 630 nm), two scatter and three fluorescence parameters - shared with Division 1.9
• MiniArrayer BioOdyssey Calligrapher with ceramic pins (BioRad) - shared with Division 1.9
• sam5 (Surface Acoustic Wave Sensor; SAW Instruments) - shared with Division 1.5

Application areas and analytes

Antibody preparations

Important application areas of our methods are:

• Environmental analysis (especially waste, surface, ground and drinking water)
• Clinical diagnostics (especially Point-of-Care Testing)

Analytes are:

• Estradiol (E2), ethinylestradiol (EE2), estrone (E1) and steroid metabolites
• Carbamazepine and metabolites
• Cetirizine
• Caffeine and other xanthines as well as their metabolites
• Coprostanol, isolithocholic acid and other sterols
• Sulfamethoxazole and other sulfonamides, including degradates/metabolites
• Laccase (an oxidase and ligninase) from the fungus Trametes versicolor
• Cardiac troponin I (cTnI)
• C-reactive protein (CRP)
• Fatty acid binding protein (FABP)

Cooperations

Often used in bioanalysis: an 8-channel pipette

Academia

• D. S. Aga; Department of Chemistry, Environmental Analytical Chemistry, University at Buffalo, The State University of New York, Buffalo, NY, USA
• V. I. Esteves, D. L. D. Lima, V. Calisto; Department of Chemistry & Center for Environmental and Marine Studies (CESAM), University of Aveiro, Aveiro, Portugal
• J. Tomé; Department of Chemistry; University of Aveiro, Aveiro, Portugal
• L.-A. Garbe; Institute of Biotechnology; Division Bioanalytics, Technische Universtität Berlin, Berlin, Germany
• B. Ondruschka, P. Bräutigam, Technical Chemistry and Environmental Chemistry; Jena University, Jena, Germany
• J. Raba, G. A. Messina, INQUISAL, Chemistry Department, Universidad Nacional de San Luis, San Luis, Argentina
• S. A. Eremin, Department of Chemistry, Division of Chemical Enzymology, Immunoassay Group, M.V. Lomonosov Moscow State University, Moscow, Russia
• M. Jekel, Department of Environmental Technology, Chair of Water Quality Control, Technische Universität Berlin, Berlin, Germany

Companies

• J. A. Schenk, Hybrotec GmbH, Potsdam
• I. Renneberg, M. Lehmann, 8sens.biognostic GmbH, Berlin
• A. Mattulat, PiCA Prüfinstitut Chemische Analytik GmbH; Berlin
• U. Dahmen-Levison, F. Mallwitz; aokin AG, Berlin
• U. Dünnbier, Berliner Wasserbetriebe (BWB)
• G. Peine, zmdb – Center for Molecular Diagnostics and Bioanalytics, Berlin

Membership

• DiagnostikNet BB – Netzwerk Diagnostik Berlin-Brandenburg e.V.

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