Molecular Bio-Engineering

The goal of the unit is the implementation of natural biological processes and systems into artificial architectures and strategies. This is achieved through the insulation of cell structures and mechanisms as well as the recombination and new orientation outside their natural environment. Transmembrane proteins could be synthesized as anchorage for extracellular functionalities and functionally expressed in cells.

An additional main focus is the generation of new immune-dominant antigens from procaryontic cDNA banks as well as work with anti-microbial peptides (AMPs). The latter offer great potential with respect to a microbial load reduction of surfaces as well as an alternative to the application of antibiotics.

  • Anti-microbial peptides for the biocidal modification of surfaces
  • Anti-microbial peptides as “capture tools” for the insulation of microorganisms
  • Nature-identical and artificial antimicrobial peptides (custom-made) for therapy and diagnostics (as alternative to antibiotics)
  • Antibody optimisation and generation of high-affinity binders
  • Procaryontic cDNA libraries
  • Detection of pathogenic microbes
  • Gentle extraction of antigens from (pathogenic) microbes
  • Construction and generation of multimeric zinc finger both technologically from genes as well as from synthetic zinc finger peptides
  • Modification of zinc fingers as DNA probes and for diagnostics
  • Modification of zinc fingers into transcription factors (synthetically and as fusion protein)
  • Zinc finger & bacteriophages as therapy (alternative to antibiotics)
  • Nucleic acid structures (self-assembly) on surfaces
  • Biological processes at/on surfaces (PCR, transcription, translation)
  • SNP detection on surfaces
  • Pathogen detection on surfaces
  • Work according to German Gene Technology Act (GenTSV) to Safety Class 2
  • Work according to the Protection Against Infection Act of Safety Class 2, with regulatory authority approval to Class 3
  • Work according to Animal Pathogen Ordinance to Safety Class 2
  • Work with obligative and facultative aerobic and anaerobic microbes
  • PCR analyses
  • C2H2 zinc finger applications (strategy, development, construction)
  • Strategy development for biochip analysis/diagnostics
  • Nucleic acid tools (stem-loop, hairpin, grids etc.)
  • Nucleic acid template design for in-vitro applications (e.g. hybridisations)
  • Amino acid templates design for in-vitro applications (e.g. binding assays)
  • All applications of classical molecular biology

Detection of Bacterial Microbes

New Immuno-Dominant Antigens

A method was developed within the scope of this project which permits the creation of new and strongly immuno-reactive proteins (antigens) from various pathogenic bacteria in a short time period and with generated prokaryotic cDNA libraries. The proteins thus obtained serve as the basis for the development of specific monoclonal antibodies and new vaccines.

New Detection Methods for Bacterial Pathogens

Bacterial pathogens with isothermal amplification analogous to PCR are detected and/or differentiated. In contrast to PCR, the multiplex-capable method requires no apparatus and is therefore for suitable for the development of new detection systems in the point-of-care sector.

Simple Quantitative Nonspecific Quick Test for Active Microbes

An ATP/NADH-based wipe test was developed in the style of a handkerchief laboratory that can ad-hoc detect potentially infectious microbes by a colour change. The process is suitable for the development of various point-of-care systems.

Antimicrobial Peptides

Insulation of Microbial Microbes

Considering a planned EU regulation for the qualification and quantification of probiotic foods, a method was developed to isolate bacterial microorganisms by means of specifically modified carrier materials made of heterogeneous mixtures, especially from yogurt. The process is based on the strong affinity of antimicrobial peptides to bacterial membranes. In connection with these investigations, complex effective principles of antimicrobial peptides could moreover be described in detail. Based on these findings, it is now possible to manufacture customized, very short, artificial and thereby non-toxic peptides (“synthicides”), compared to peptides that occur in nature.

Antimicrobial Surfaces

This project generated biocidic surfaces on the basis of peptides that reduce microbial load. Since bacteria can form resistances against the short-chained peptides only with great difficulty, this is an effective alternative to the use of antibiotics and can help to brake growing antibiotic resistances.

  • PCR technologies (RT, real-time, quantitative, on-chip, in-situ, gradient)
  • Laboratory equipment for work with genetically engineered organisms, infectious microbes and animal pathogens (S1/S2, cell culture, yeast laboratory, bacteria laboratory)
  • RNA laboratory
  • Controllable fumigated glovebox
  • Electrophoresis (Agarose, Acrylamid)
  • UV-vis spectrophotometer
  • Gel Imager
  • Centrifuges (cooled, high speed, large and small volumes, ultra-)
  • Incubators (pro- and eukaryotic)
  • Hybridisation equipment (some fully-automatic)

Publications

  • Danckert L, Hoppe S, Bier FF, von Nickisch-Rosenegk M. Rapid identification of novel antigens of Salmonella Enteritidis by microarray-based immunoscreening. Microchimica Acta 02/2014; 3.43 Impact Factor.
  • Herbel S, von Nickisch-Rosenegk M, Kuhn M, Murugaiyan J, Wieler LH, Guenther S. Specific TaqMan Probes for the Identification and Quantification of Lactobacilli in Pharmaceuticals. Journal of Probiotics & Health. 02/2014; 2(1).
  • Hoppe S, Bier FF, von Nickisch-Rosenegk M. Identification of antigenic proteins of the nosocomial pathogen Klebsiella pneumoniae. PLoS One 9(10): e110703. Doi dx.doi.org/10.1371/journal.pone.0110703.
  • Kersting S, Rausch V, Bier FF, von Nickisch-Rosenegk M. Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis. Malaria Journal 03/2014; 13(1):99.
  • Kersting S, Rausch V, Bier FF, von Nickisch-Rosenegk M. Multiplex isothermal solid-phase recombinase polymerase amplification for the specific and fast DNA-based detection of three bacterial pathogens. Microchimica Acta 02/2014.
  • Rapsch K, Bier FF, von Nickisch-Rosenegk M. Rational Design of Artificial Beta-Strand-Forming Antimicrobial Peptides with Biocompatible Properties. Mol Pharm. 2014 Oct 6;11(10):3492-502. doi: 10.1021/mp500271c. Epub 2014 Sep 22. PubMed PMID: 25192319.
  • Herbel SR, Lauzat B, von Nickisch-Rosenegk M, Kuhn M, Murugaiyan J, Wieler LH, Guenther S. Species-specific quantification of probiotic lactobacilli in yoghurt by quantitative real time PCR. Journal of Applied Microbiology 09/2013.
  • Hoppe S, Bier FF, von Nickisch-Rosenegk M. Rapid Identification of Novel Immunodominant Proteins and Characterization of a Specific Linear Epitope of Campylobacter jejuni. PLoS ONE 01/2013; 8(5):e65837.
  • Rapsch K, Bier FF, Tadros M, von Nickisch-Rosenegk M. Identification of antimicrobial peptides and immobilization strategy suitable for a covalent surface coating with biocompatible properties. Bioconjugate Chemistry 12/2013.
  • Hoppe S, Bier FF, von Nickisch-Rosenegk M. Microarray-based method for screening of immunogenic proteins from bacteria. Journal of Nanobiotechnology 03/2012; 10:12.
  • von Nickisch-Rosenegk M, Teschke T, Bier FF. Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells. Journal of Nanobiotechnology 01/2012; 10:1.
  • Andresen D, von Nickisch-Rosenegk M, Bier FF. Helicase-dependent amplification: use in OnChip amplification and potential for point-of-care diagnostics. Expert Review of Molecular Diagnostics 10/2009; 9(7):645-50.
  • Andresen D, von Nickisch-Rosenegk M, Bier FF. Helicase dependent OnChip-amplification and its use in multiplex pathogen detection. Clinica chimica acta; international journal of clinical chemistry 04/2009; 403(1-2):244-8.
  • Guenther S, Landgraf M, von Nickisch-Rosenegk M, Kuhn M, Wieler LH, Bier F, Schierack P. "PorkChip"-Detektionssystem zum schnellen und parallelen Nachweis von Zoonoseerregern aus dem Schwein innerhalb der Lebensmittelkette. RFL. 12/2008; 12.
  • Guenther S, Nöckler K, von Nickisch-Rosenegk M, Landgraf M, Ewers C, Wieler LH, Schierack P. Detection of Trichinella spiralis, T. britovi and T. pseudospiralis in muscle tissue with real-time PCR. Journal of Microbiological Methods 10/2008; 75(2):287-92.
  • von Nickisch-Rosenegk M, Marschan X, Andresen D, Bier FF. Reverse transcription-polymerase chain reaction on a microarray: the integrating concept of "active arrays". Analytical and Bioanalytical Chemistry 08/2008; 391(5):1671-8.
  • Bier FF, von Nickisch-Rosenegk M, Ehrentreich-Förster E, Reiss E, Henkel J, Strehlow R, Andresen D. DNA microarrays. Advances in Biochemical Engineering/Biotechnology 02/2008; 109:433-53.
  • von Nickisch-Rosenegk M, Ehrentreich-Förster E, Strehlow R, Christmann A, Bier FF. Chemically synthesized zinc finger molecules as nano-addressable probes for double-stranded DNAs. Journal of Nanobiotechnology 07/2005; 3:5.
  • Steffen J, von Nickisch-Rosenegk M, Bier FF. In vitro transcription of a whole gene on a surface-coupled template. Lab on a Chip 07/2005; 5(6):665-8.
  • von Nickisch-Rosenegk M, Marschan X, Andresen D, Abraham A, Heise C, Bier FF. On-chip PCR amplification of very long templates using immobilized primers on glassy surfaces. Biosensors and Bioelectronics 03/2005; 20(8):1491-8.

Patents

  • von Nickisch-Rosenegk M. Genetically modified bacteriophages, in particular for combating pathogenic prokaryotes or their pathological effect, and its use and production. WO 2008/086881 A1. 24 July 2008.
  • von Nickisch-Rosenegk M, Bier F, Andresen H. Means and methods for producing zinc fingers and concatemers thereof. EP 2 130 836 B1. 13. Feb. 2013.