@article{HimmelVanderVenStoeckleinetal.2003,
  author    = {Himmel, Mirko and VanderVen, Peter F. M. and St{\"o}cklein, Walter F. M. and F{\"u}rst, Dieter Oswald},
  title     = {The limits of promiscuity : isoform-specific dimerization of filamins},
  year      = {2003},
  language  = {en}
}
@article{LisdatUtepbergenovHaseloffetal.2001,
  author    = {Lisdat, Fred and Utepbergenov, D. and Haseloff, R. F. and Blasig, Ingolf E. and St{\"o}cklein, Walter F. M. and Scheller, Frieder W. and Brigelius-Floh{\´e}, Regina},
  title     = {An optical method for the detection of oxidative stress using protein-RNA interaction},
  year      = {2001},
  language  = {en}
}
@article{StoeckleinRohdeScharteetal.2000,
  author    = {St{\"o}cklein, Walter F. M. and Rohde, M. and Scharte, Gudrun and Behrsing, Olaf and Warsinke, Axel and Micheel, Burkhard and Scheller, Frieder W.},
  title     = {Sensitive detection of triazine and phenylurea pesticides in pure organic solvent by enzyme linked immunsorbent assay (ELISA): stabilities, solubilities and sensitives},
  year      = {2000},
  language  = {en}
}
@article{NitscheKurthDunkhorstetal.2007,
  author    = {Nitsche, Andreas and Kurth, Andreas and Dunkhorst, Anna and P{\"a}nke, Oliver and Sielaff, Hendrik and Junge, Wolfgang and Muth, Doreen and Scheller, Frieder W. and St{\"o}cklein, Walter F. M. and Pauli, Georg and Kage, Andreas},
  title     = {One-step selection of vaccinia virus binding DNA-aptamers by MonoLEX},
  doi       = {10.1186/1472-6750-7-48},
  year      = {2007},
  language  = {en}
}
@article{NeumannSchulteJuenemannetal.2006,
  author    = {Neumann, Meina and Schulte, Marc and J{\"u}nemann, Nora and St{\"o}cklein, Walter F. M. and Leimk{\"u}hler, Silke},
  title     = {Rhodobacter capsulatus XdhC is involved in molybdenum cofactor binding and insertion into xanthine dehydrogenase},
  doi       = {10.1074/jbc.M601617200},
  year      = {2006},
  abstract  = {Rhodobacter capsulatus xanthine dehydrogenase (XDH) is a cytoplasmic enzyme with an (alpha beta) 2 heterodimeric structure that is highly identical to homodimeric eukaryotic xanthine oxidoreductases. The crystal structure revealed that the molybdenum cofactor (Moco) is deeply buried within the protein. A protein involved in Moco insertion and XDH maturation has been identified, which was designated XdhC. XdhC was shown to be essential for the production of active XDH but is not a subunit of the purified enzyme. Here we describe the purification of XdhC and the detailed characterization of its role for XDH maturation. We could show that XdhC binds Moco in stoichiometric amounts, which subsequently can be inserted into Moco-free apo-XDH. A specific interaction between XdhC and XdhB was identified. We show that XdhC is required for the stabilization of the sulfurated form of Moco present in enzymes of the xanthine oxidase family. Our findings imply that enzyme-specific proteins exist for the biogenesis of molybdoenzymes, coordinating Moco binding and insertion into their respective target proteins. So far, the requirement of such proteins for molybdoenzyme maturation has been described only for prokaryotes},
  language  = {en}
}
@article{BeissenhirtzSchellerStoeckleinetal.2004,
  author    = {Beissenhirtz, Moritz Karl and Scheller, Frieder W. and St{\"o}cklein, Walter F. M. and Kurth, D. and M{\"o}hwald, Helmuth and Lisdat, Fred},
  title     = {Electroactive cytochrome c multilayers within a polyelectrolyte assembly},
  year      = {2004},
  language  = {en}
}
@article{Stoecklein2006,
  author    = {St{\"o}cklein, Walter F. M.},
  title     = {Molecule-detective},
  year      = {2006},
  abstract  = {Biosensors are analytical devices incorporating biological material (receptor) intimately associated with or integrated within a physicochernical transducer. Advantages are the high selectivity for analyte detection. Examples given comprise the very successful commercial blood glucose biosensors made for the self-control by the diabetic patients. Other biosensors are part of an analytic system, including the sensor chips Of surface plasmon resonance or interferometry based devices, piezoelectric or reflectometric sensors capable of direct measurement of mass changes, and thermometric and other reagentless sensors. The development of nanotubes-based devices allows for significant enhancment of the signal-to-noise ratio of the biosensors. A milestone on the way towards miniaturization and parallelization of biosensors is the recently developed and prize-winning electronic DNA chip},
  language  = {en}
}
@article{LiuWollenbergerHalameketal.2005,
  author    = {Liu, Songqin and Wollenberger, Ursula and Halamek, Jan and Leupold, Eik and St{\"o}cklein, Walter F. M. and Warsinke, Axel and Scheller, Frieder W.},
  title     = {Affinity interaction betwen phenylboronic acid-carrying self-assembled monolayers and FAD or HRP},
  year      = {2005},
  abstract  = {A method is provided for the recognition of glycated molecules based on their binding affinities to boronate- carrying monolayers. The affinity interaction of flavin adenine dinucleotide (FAD) and horseradish peroxidase (HRP) with phenylboronic acid monolayers on gold was investigated by using voltammetric and microgravimetric methods. Conjugates of 3-aminopherrylboronic acid and 3,3'-dithiodipropionic acid di(N-hydroxysuccinimide ester) or 11-mercaptoundecanoic acid were prepared and self-assembled on gold surfaces to generate monolayers. FAD is bound to this modified sur-face and recognized by a pair of redox peaks with a formal potential of -0.433 V in a 0.1 m phosphate buffer solution, pH 6.5. Upon addition of a sugar to the buffer, the bound FAD could be replaced, indicating that the binding is reversible. Voltammetric, mass measurements, and photometric activity assays show that the HRP can also be bound to the interface. This binding is reversible, and HRP can be replaced by sorbitol or removed in acidic solution. The effects of pH, incubation time, and concentration of H2O2 were studied by comparing the catalytic reduction of H2O2 in the presence of the electron-donor thionine. The catalytic current of the HRP-loaded electrode was proportional to HRP concentrations in the incubation solution in the range between 5 mu g mL(-1) and 0.4 mg mL(-1) with a linear slope of 3.34 mu A mL mg(-1) and a correlation coefficient of 0.9945},
  language  = {en}
}
@article{KleuserStoeckleinPieperFuerstetal.2004,
  author    = {Kleuser, U. and St{\"o}cklein, Walter F. M. and Pieper-F{\"u}rst, U. and Scheller, Frieder W.},
  title     = {Partikelverst{\"a}rkte Oberfl{\"a}chenplasmonresonanz f{\"u}r die Quantifizierung von Matrix Metalloproteinase-2},
  year      = {2004},
  language  = {de}
}
@article{PieperFuerstKleuserStoeckleinetal.2004,
  author    = {Pieper-F{\"u}rst, U. and Kleuser, U. and St{\"o}cklein, Walter F. M. and Warsinke, Axel and Scheller, Frieder W.},
  title     = {Detection of subicomolar concentrations of human matrix metalloproteinase-2 by an optical biosensor},
  year      = {2004},
  abstract  = {We describe in this paper the development of a one-step sandwich assay for the highly sensitive and fast detection of human matrix metalloproteinase (MMP)-2 (EC 3.4.24.24), using surface plasmon resonance (SPR). For the assay, two ligands were selected: monoclonal anti-MMP-2 antibody Ab-2 and the tissue inhibitor of metalloproteinases (TIMP)-2. They were chosen on the basis of (1) their affinities to MMP-2, (2) the efficiency of immobilization to the sensor chip, (3) the efficiency of adsorption to colloidal gold, and (4) the stability of these protein-coated gold particles. The assay included mixing of MMP-2 with antibody Ab-2 adsorbed to colloidal gold with a diameter of about 20 rim and injection into the flowcell of the SPR instrument containing immobilized TIMP-2. By using colloidal gold particles an amplification factor of 114 and a detection limit of 0.5 pM for MMP-2 were obtained. The precision of the assay was high even at low analyte concentrations, the standard deviation being 8.3\% for five determinations of 1 pM MMP- 2. No significant binding was observed with the structurally related MMP-9. The assay is far more sensitive and faster than commonly used methods for MMP-2 detection. As TIMP-bound MMP-2 is not detected by this method, the assay can be applied for measuring free MMP-2, reflecting the imbalance of free and inhibitor-bound enzyme in various pathological situations. (C) 2004 Elsevier Inc. All rights reserved},
  language  = {en}
}
@article{ChenStoeckleinSchelleretal.2003,
  author    = {Chen, Jian and St{\"o}cklein, Walter F. M. and Scheller, Frieder W. and Wollenberger, Ursula},
  title     = {Electrochemical determination of human hemoglobin by using ferrocene carboxylic acid modified carbon powder microelectrode},
  year      = {2003},
  language  = {en}
}
@article{NiessnerBroekaertEinaxetal.2002,
  author    = {Nießner, Reinhard and Broekaert, J. and Einax, J. W. and Scheller, Frieder W. and St{\"o}cklein, Walter F. M.},
  title     = {Trendbericht analytische Biochemie 2000/2001},
  year      = {2002},
  language  = {de}
}
@article{SassStoeckleinKlevesathetal.2019,
  author    = {Sass, Stephan and St{\"o}cklein, Walter F. M. and Klevesath, Anja and Hurpin, Jeanne and Menger, Marcus and Hille, Carsten},
  title     = {Binding affinity data of DNA aptamers for therapeutic anthracyclines from microscale thermophoresis and surface plasmon resonance spectroscopy},
  series = {The analyst : the analytical journal of the Royal Society of Chemistry},
  volume    = {144},
  journal   = {The analyst : the analytical journal of the Royal Society of Chemistry},
  number    = {20},
  publisher = {Royal Society of Chemistry},
  address   = {Cambridge},
  issn      = {0003-2654},
  doi       = {10.1039/c9an01247h},
  pages     = {6064 -- 6073},
  year      = {2019},
  abstract  = {Anthracyclines like daunorubicin (DRN) and doxorubicin (DOX) play an undisputed key role in cancer treatment, but their chronic administration can cause severe side effects. For precise anthracycline analytical systems, aptamers are preferable recognition elements. Here, we describe the detailed characterisation of a single-stranded DNA aptamer DRN-10 and its truncated versions for DOX and DRN detection. Binding affinities were determined from surface plasmon resonance (SPR) and microscale thermophoresis (MST) and combined with conformational data from circular dichroism (CD). Both aptamers displayed similar nanomolar binding affinities to DRN and DOX, even though their rate constants differed as shown by SPR recordings. SPR kinetic data unravelled a two-state reaction model including a 1 : 1 binding and a subsequent conformational change of the binding complex. This model was supported by CD spectra. In addition, the dissociation constants determined with MST were always lower than that from SPR, and especially for the truncated aptamer they differed by two orders of magnitude. This most probably reflects the methodological difference, namely labelling for MST vs. immobilisation for SPR. From CD recordings, we suggested a specific G-quadruplex as structural basis for anthracycline binding. We concluded that the aptamer DRN-10 is a promising recognition element for anthracycline detection systems and further selected aptamers can be also characterised with the combined methodological approach presented here.},
  language  = {en}
}
@article{StoeckleinWarsinkeScheller1997,
  author    = {St{\"o}cklein, Walter F. M. and Warsinke, Axel and Scheller, Frieder W.},
  title     = {Organic solvent modified enzyme-liked immunoassay for the detection of triazine herbicides},
  year      = {1997},
  language  = {en}
}
@article{StoeckleinScheller1997,
  author    = {St{\"o}cklein, Walter F. M. and Scheller, Frieder W.},
  title     = {Enzymes and antibodies in organic media : analytical applications},
  year      = {1997},
  language  = {en}
}
@article{KramerKeitelWinkleretal.1997,
  author    = {Kramer, A. and Keitel, T. and Winkler, K. and St{\"o}cklein, Walter F. M. and H{\"u}hne, Wolfgang and Schneider-Mergener, Jens},
  title     = {Molecular basis for the binding promiscuity of an anti-P24 (HIV-1) monoclonal antibody},
  year      = {1997},
  language  = {en}
}
@article{WollenbergerDrungilieneStoeckleinetal.1996,
  author    = {Wollenberger, Ursula and Drungiliene, A. and St{\"o}cklein, Walter F. M. and Kulys, J. and Scheller, Frieder W.},
  title     = {Direct electrocatalytic determination of dissolved peroxidases},
  year      = {1996},
  language  = {en}
}
@article{StoeckleinSchellerAbuknesha1995,
  author    = {St{\"o}cklein, Walter F. M. and Scheller, Frieder W. and Abuknesha, Rhamadan},
  title     = {Effects of organic solvents on semicontinuous immunochemical detection of coumarin derivatives},
  year      = {1995},
  language  = {en}
}
@article{StoeckleinScheller1995,
  author    = {St{\"o}cklein, Walter F. M. and Scheller, Frieder W.},
  title     = {Einsatz von Enzymen und Antik{\"o}rpern in organischen L{\"o}sungsmitteln},
  year      = {1995},
  language  = {de}
}
@article{StoeckleinWarsinkeMicheeletal.1998,
  author    = {St{\"o}cklein, Walter F. M. and Warsinke, Axel and Micheel, Burkhard and H{\"o}hne, Wolfgang and Woller, Jochen and Kempter, Gerhard and Scheller, Frieder W.},
  title     = {Characterization of a monoclonal antibody and its Fab fragment against diphenylurea hapten with BIA},
  isbn      = {3-8154-3540-4},
  year      = {1998},
  language  = {en}
}
@article{StoeckleinWarsinkeMicheeletal.1997,
  author    = {St{\"o}cklein, Walter F. M. and Warsinke, Axel and Micheel, Burkhard and H{\"o}hne, Wolfgang and Woller, Jochen and Kempter, Gerhard and Scheller, Frieder W.},
  title     = {Detection of diphenylurea derivatives with biospecific interaction analysis (BIA) : Kinetic investigations},
  year      = {1997},
  language  = {en}
}
@article{JinBernhardtStoeckleinetal.1998,
  author    = {Jin, Wen and Bernhardt, Rita and St{\"o}cklein, Walter F. M. and Scheller, Frieder W.},
  title     = {Direct electron transfer of adrenodoxin-a [2Fe-2S] protein-- and its mutants on modified gold electrode},
  year      = {1998},
  language  = {en}
}
@article{StoeckleinScheller1996,
  author    = {St{\"o}cklein, Walter F. M. and Scheller, Frieder W.},
  title     = {Laccase : a marker enzyme for solvent modified immunoassays},
  year      = {1996},
  language  = {en}
}
@article{HalamekWollenbergerStoeckleinetal.2007,
  author    = {Hal{\´a}mek, Jan and Wollenberger, Ursula and St{\"o}cklein, Walter F. M. and Scheller, Frieder W.},
  title     = {Development of a biosensor for glycated hemoglobin},
  issn      = {0013-4686},
  doi       = {10.1016/j.electacta.2007.03.059},
  year      = {2007},
  abstract  = {The development of an electrochemical piezoelectric sensor for the detection of glycated hemoglobin is presented. The total hemoglobin (Hb) content is monitored with a mass-sensitive quartz crystal modified with surfactants, and the glycated fraction of the immobilized Hb is determined by subsequent voltarnmetric measurement of the coupled ferroceneboronic acid. Different modifications of the sensor were tested for their hemoglobin binding ability. Deoxycholate (DOCA) was found to be the most suitable among the examined modifiers. Piezoelectric quartz crystals with gold electrodes were modified with DOCA by covalent binding to a pre-formatted 4-aminothiophenol monolayer. The properties of the Hb binding to DOCA and the pH effect on this interaction were studied. In the proposed assay for glycated hemoglobin at first an Hb sample is incubated with ferroceneboronic acid (FcBA), which binds to the fructosyl residue of the glycated Hb. Then this preincubated Hb sample is allowed to interact with the DOCA-modified piezoelectric quartz crystal. The binding is monitored by quartz crystal nanobalance QCN). The amount of FcBA present on the sensor surface is determined by square wave voltammetry. The binding of FcBA results in well-defined peaks with an EO' of +200 mV versus Ag/AgC1 (1 M KC1). The peak height depends on the degree of glycated Hb in the sample ranging from 0\% to 20\% of total Hb. The regeneration of the sensing surface is achieved by pepsin digestion of the deposited Hb. Thus the sensor can be re-used more than 30 times.},
  language  = {en}
}
@article{HalamekWollenbergerStoeckleinetal.2007,
  author    = {Hal{\´a}mek, Jan and Wollenberger, Ursula and St{\"o}cklein, Walter F. M. and Warsinke, Axel and Scheller, Frieder W.},
  title     = {Signal amplification in immunoassays using labeling via boronic acid binding to the sugar moiety of immunoglobulin G : proof of concept for glycated hemoglobin},
  issn      = {0003-2719},
  doi       = {10.1080/00032710701327096},
  year      = {2007},
  abstract  = {A novel electrochemical immunoassay based on the multiple affinity labeling of the indicator antibody with an electro-active tag is presented. The concept is illustrated for the determination of the glycated hemoglobin HbA1c in hemoglobin samples. Hemoglobin is adsorbed to the surfactant-modified surface of a piezoelectric quartz crystal. Whereas the quartz crystal nanobalance is used to validate the total Hb binding, the HbA1c on the sensor surface is recognized by an antibody and quantified electrochemically after the sugar moieties of the antibody have been labeled in-situ with ferroceneboronic acid. The sensitivity of this sensor is about threefold higher than the sensitivity of a hemoglobin sensor, where the ferroceneboronic acid is bound directly to HbA1c.},
  language  = {en}
}
@article{StoeckleinWarsinkeMicheeletal.1998,
  author    = {St{\"o}cklein, Walter F. M. and Warsinke, Axel and Micheel, Burkhard and Kempter, Gerhard and H{\"o}hne, Wolfgang and Scheller, Frieder W.},
  title     = {Diphenylurea hapten sensing with a monoclonal antibody and its Fab fragment : kinetic and thermodynamic investigations},
  year      = {1998},
  language  = {en}
}
@article{WollenbergerJinBernhardtetal.1998,
  author    = {Wollenberger, Ursula and Jin, Wen and Bernhardt, Rita and Lehmann, Claudia and St{\"o}cklein, Walter F. M. and Brigelius-Floh{\´e}, Regina and Scheller, Frieder W.},
  title     = {Funktionalisierung von Elektroden f{\"u}r den direkten heterogenen Elektrotransfer},
  year      = {1998},
  language  = {de}
}
@article{StoellnerStoeckleinSchelleretal.2002,
  author    = {St{\"o}llner, Daniela and St{\"o}cklein, Walter F. M. and Scheller, Frieder W. and Warsinke, Axel},
  title     = {Membrane-immobilized haptoglobin as affinity matrix for a hemoglobin-A1c-immunosensor},
  year      = {2002},
  language  = {en}
}
@article{Stoecklein2000,
  author    = {St{\"o}cklein, Walter F. M.},
  title     = {Biosensoren f{\"u}r die direkte vor-Ort {\"U}berwachung von Umwelt-Schadstoffen},
  year      = {2000},
  language  = {de}
}
@article{StoeckleinBehrsingScharteetal.2000,
  author    = {St{\"o}cklein, Walter F. M. and Behrsing, Olaf and Scharte, Gudrun and Micheel, Burkhard and Benkert, Alexander and Sch{\"o}ssler, W. and Warsinke, Axel and Scheller, Frieder W.},
  title     = {Enzyme kinetic assays with surface plasmon resonance (BIAcore) based on competition between enzyme and creatinine antibody},
  year      = {2000},
  language  = {en}
}
@article{LisdatGeStoeckleinetal.2000,
  author    = {Lisdat, Fred and Ge, Bixia and St{\"o}cklein, Walter F. M. and Scheller, Frieder W. and Meyer, T.},
  title     = {Electrochemical behaviour and nitric oxides interaction of immobilised cytochrome c from Rhodocyclus gelatinosus},
  year      = {2000},
  language  = {en}
}
@article{StoeckleinMakowerBieretal.1997,
  author    = {St{\"o}cklein, Walter F. M. and Makower, Alexander and Bier, Frank Fabian and Scheller, Frieder W.},
  title     = {Enzyme sensors and enzyme amplifification systems},
  year      = {1997},
  language  = {en}
}
@article{HovestaedtMemczakPleineretal.2014,
  author    = {Hovestaedt, Marc and Memczak, Henry and Pleiner, Dennis and Zhang, Xin and Rappich, Joerg and Bier, Frank Fabian and St{\"o}cklein, Walter F. M.},
  title     = {Characterization of a new maleimido functionalization of gold for surface plasmon resonance spectroscopy},
  series = {Journal of molecular recognition : an international journal devoted to research on specific molecular recognition in chemistry, biology, biotechnology and medicine},
  volume    = {27},
  journal   = {Journal of molecular recognition : an international journal devoted to research on specific molecular recognition in chemistry, biology, biotechnology and medicine},
  number    = {12},
  publisher = {Wiley-Blackwell},
  address   = {Hoboken},
  issn      = {0952-3499},
  doi       = {10.1002/jmr.2396},
  pages     = {707 -- 713},
  year      = {2014},
  abstract  = {Para-maleimidophenyl (p-MP) modified gold surfaces have been prepared by one-step electrochemical deposition and used in surface plasmon resonance (SPR) studies. Therefore, a FITC mimotope peptide (MP1, 12 aa), a human mucin 1 epitope peptide (MUC, 9 aa) and a protein with their specific antibodies were used as model systems. The peptides were modified with an N-terminal cysteine for covalent and directed coupling to the maleimido functionalized surface by means of Michael addition. The coupling yield of the peptide, the binding characteristics of antibody and the unspecific adsorption of the analytes were investigated. The results expand the spectrum of biosensors usable with p-MP by widely used SPR and support its potential to be versatile for several electrochemical and optical biosensors. This allows the combination of an electrochemical and optical read-out for a broad variety of biomolecular interactions on the same chip. Copyright (c) 2014 John Wiley \& Sons, Ltd.},
  language  = {en}
}
@misc{MemczakLausterKaretal.2016,
  author    = {Memczak, Henry and Lauster, Daniel and Kar, Parimal and Di Lella, Santiago and Volkmer, Rudolf and Knecht, Volker and Herrmann, Andreas and Ehrentreich-F{\"o}rster, Eva and Bier, Frank Fabian and St{\"o}cklein, Walter F. M.},
  title     = {Anti-hemagglutinin antibody derived lead peptides for inhibitors of influenza virus binding},
  series = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe},
  journal   = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe},
  number    = {536},
  issn      = {1866-8372},
  doi       = {10.25932/publishup-41087},
  url       = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-410872},
  pages     = {24},
  year      = {2016},
  abstract  = {Antibodies against spike proteins of influenza are used as a tool for characterization of viruses and therapeutic approaches. However, development, production and quality control of antibodies is expensive and time consuming. To circumvent these difficulties, three peptides were derived from complementarity determining regions of an antibody heavy chain against influenza A spike glycoprotein. Their binding properties were studied experimentally, and by molecular dynamics simulations. Two peptide candidates showed binding to influenza A/Aichi/2/68 H3N2. One of them, termed PeB, with the highest affinity prevented binding to and infection of target cells in the micromolar region without any cytotoxic effect. PeB matches best the conserved receptor binding site of hemagglutinin. PeB bound also to other medical relevant influenza strains, such as human-pathogenic A/California/7/2009 H1N1, and avian-pathogenic A/MuteSwan/Rostock/R901/2006 H7N1. Strategies to improve the affinity and to adapt specificity are discussed and exemplified by a double amino acid substituted peptide, obtained by substitutional analysis. The peptides and their derivatives are of great potential for drug development as well as biosensing.},
  language  = {en}
}
@inproceedings{MemczakLausterHerrmannetal.2013,
  author    = {Memczak, Henry and Lauster, Daniel and Herrmann, Andreas and St{\"o}cklein, Walter F. M. and Bier, Frank Fabian},
  title     = {Novel hemagglutinin-binding peptides for biosensing and inhibition of Influenza Viruses},
  series = {Biopolymers},
  volume    = {100},
  booktitle = {Biopolymers},
  number    = {3},
  publisher = {Wiley-Blackwell},
  address   = {Hoboken},
  issn      = {0006-3525},
  pages     = {255 -- 255},
  year      = {2013},
  language  = {en}
}
@article{StechMerkSchenketal.2012,
  author    = {Stech, Marlitt and Merk, Helmut and Schenk, J{\"o}rg A. and St{\"o}cklein, Walter F. M. and W{\"u}stenhagen, Doreen Anja and Micheel, Burkhard and Duschl, Claus and Bier, Frank Fabian and Kubick, Stefan},
  title     = {Production of functional antibody fragments in a vesicle-based eukaryotic cell-free translation system},
  series = {Journal of biotechnology},
  volume    = {164},
  journal   = {Journal of biotechnology},
  number    = {2},
  publisher = {Elsevier},
  address   = {Amsterdam},
  issn      = {0168-1656},
  doi       = {10.1016/j.jbiotec.2012.08.020},
  pages     = {220 -- 231},
  year      = {2012},
  abstract  = {Cell-free protein synthesis is of increasing interest for the rapid and high-throughput synthesis of many proteins, in particular also antibody fragments. In this study, we present a novel strategy for the production of single chain antibody fragments (scFv) in a eukaryotic in vitro translation system. This strategy comprises the cell-free expression, isolation and label-free interaction analysis of a model antibody fragment synthesized in two differently prepared insect cell lysates. These lysates contain translocationally active microsomal structures derived from the endoplasmic reticulum (ER), allowing for posttranslational modifications of cell-free synthesized proteins. Both types of these insect cell lysates enable the synthesis and translocation of scFv into ER-derived vesicles. However, only the one that has a specifically adapted redox potential yields functional active antibody fragments. We have developed a new methodology for the isolation of functional target proteins based on the translocation of cell-free produced scFv into microsomal structures and subsequent collection of protein-enriched vesicles. Antibody fragments that have been released from these vesicles are shown to be well suited for label-free binding studies. Altogether, these results show the potential of insect cell lysates for the production, purification and selection of antibody fragments in an easy-to-handle and time-saving manner.},
  language  = {en}
}
@article{EisoldSellrieSchenketal.2015,
  author    = {Eisold, Ursula and Sellrie, Frank and Schenk, J{\"o}rg A. and Lenz, Christine and St{\"o}cklein, Walter F. M. and Kumke, Michael Uwe},
  title     = {Bright or dark immune complexes of anti-TAMRA antibodies for adapted fluorescence-based bioanalysis},
  series = {Analytical \& bioanalytical chemistry},
  volume    = {407},
  journal   = {Analytical \& bioanalytical chemistry},
  number    = {12},
  publisher = {Springer},
  address   = {Heidelberg},
  issn      = {1618-2642},
  doi       = {10.1007/s00216-015-8538-0},
  pages     = {3313 -- 3323},
  year      = {2015},
  abstract  = {Fluorescence labels, for example fluorescein or rhodamin derivatives, are widely used in bioanalysis applications including lateral-flow assays, PCR, and fluorescence microscopy. Depending on the layout of the particular application, fluorescence quenching or enhancement may be desired as the detection principle. Especially for multiplexed applications or high-brightness requirements, a tunable fluorescence probe can be beneficial. The alterations in the photophysics of rhodamine derivatives upon binding to two different anti-TAMRA antibodies were investigated by absorption and fluorescence-spectroscopy techniques, especially determining the fluorescence decay time and steady-state and time-resolved fluorescence anisotropy. Two monoclonal anti-TAMRA antibodies were generated by the hybridoma technique. Although surface-plasmon-resonance measurements clearly proved the high affinity of both antibodies towards 5-TAMRA, the observed effects on the fluorescence of rhodamine derivatives were very different. Depending on the anti-TAMRA antibody either a strong fluorescence quenching (G71-DC7) or a distinct fluorescence enhancement (G71-BE11) upon formation of the immune complex was observed. Additional rhodamine derivatives were used to gain further information on the binding interaction. The data reveal that such haptens as 5-TAMRA could generate different paratopes with equal binding affinities but different binding interactions, which provide the opportunity to adapt bioanalysis methods including immunoassays for optimized detection principles for the same hapten depending on the specific requirements.},
  language  = {en}
}
@article{SchenkSellrieBoettgeretal.2007,
  author    = {Schenk, J{\"o}rg A. and Sellrie, Frank and B{\"o}ttger, Volker and Micheel, Burkhard and St{\"o}cklein, Walter F. M.},
  title     = {Generation and application of a fluorescein-specific single chain antibody},
  year      = {2007},
  abstract  = {A recombinant single chain antibody fragment (designated scDE1) of the murine monoclonal anti-fluorescein antibody B13-DE1 was generated using the original hybridoma cells as source for the variable antibody heavy and light chain (VH and VL) genes. After cloning the variable genes into a phage vector a functional antibody fragment was selected by phage display panning. Recombinant antibody could be expressed as phage antibody and as soluble single chain antibody in Escherichia coli. High yield of scDE1 could also be detected in bacterial culture supernatant. The scDE1 showed the same binding specificity as the parental monoclonal antibody, i.e. it bound fluorescein, fluorescein derivatives and a fluorescein peptide mimotope. Surface plasmon resonance revealed a K(D) of 19 nM for the scDE1 compared to 0.7 nM for the monoclonal antibody. The isolated soluble scDE1 could easily be conjugated to horseradish peroxidase which allowed the use of the conjugate as universal indicator for the detection of fluorescein-labelled proteins in different immunoassays. Detection of hCG in urine was performed as a model system using scDE1. In addition to E. coli the scFv genes could also be transferred and expressed in eukaryotic cells. Finally, we generated HEK293 cells expressing the scDE1 at the cell surface.},
  language  = {en}
}