Bio

Professional Education


  • Dr., Max Planck Institute for Molecular Genetics and FU Berlin (2012)
  • Diplom, Freie Universitat Berlin (2008)

Stanford Advisors


Publications

Journal Articles


  • Onset of Immune Senescence Defined by Unbiased Pyrosequencing of Human Immunoglobulin mRNA Repertoires PLOS ONE Rubelt, F., Sievert, V., Knaust, F., Diener, C., Lim, T. S., Skriner, K., Klipp, E., Reinhardt, R., Lehrach, H., Konthur, Z. 2012; 7 (11)

    Abstract

    The immune system protects us from foreign substances or pathogens by generating specific antibodies. The variety of immunoglobulin (Ig) paratopes for antigen recognition is a result of the V(D)J rearrangement mechanism, while a fast and efficient immune response is mediated by specific immunoglobulin isotypes obtained through class switch recombination (CSR). To get a better understanding on how antibody-based immune protection works and how it changes with age, the interdependency between these two parameters need to be addressed. Here, we have performed an in depth analysis of antibody repertoires of 14 healthy donors representing different gender and age groups. For this task, we developed a unique pyrosequencing approach, which is able to monitor the expression levels of all immunoglobulin V(D)J recombinations of all isotypes including subtypes in an unbiased and quantitative manner. Our results show that donors have individual immunoglobulin repertoires and cannot be clustered according to V(D)J recombination patterns, neither by age nor gender. However, after incorporating isotype-specific analysis and considering CSR information into hierarchical clustering the situation changes. For the first time the donors cluster according to age and separate into young adults and elderly donors (>50). As a direct consequence, this clustering defines the onset of immune senescence at the age of fifty and beyond. The observed age-dependent reduction of CSR ability proposes a feasible explanation why reduced efficacy of vaccination is seen in the elderly and implies that novel vaccine strategies for the elderly should include the "Golden Agers".

    View details for DOI 10.1371/journal.pone.0049774

    View details for Web of Science ID 000312376100028

    View details for PubMedID 23226220

  • A streamlined protocol for emulsion polymerase chain reaction and subsequent purification ANALYTICAL BIOCHEMISTRY Schuetze, T., Rubelt, F., Repkow, J., Greiner, N., Erdmann, V. A., Lehrach, H., Konthur, Z., Gloekler, J. 2011; 410 (1): 155-157

    Abstract

    Compartmentalization of polymerase chain reaction (PCR) reduces artifacts, especially when complex libraries are amplified. It allows clonal amplification of templates from complex mixtures in a bias-free manner. Here we describe a rapid, straightforward, and easy protocol for PCR in a water-in-oil emulsion (ePCR) including sample recovery by DNA purification. Furthermore, no special laboratory equipment is needed and inexpensive components are used. Therefore, our flexible protocol allows ePCR to be readily implemented in daily routine experiments for a broad range of applications.

    View details for DOI 10.1016/j.ab.2010.11.029

    View details for Web of Science ID 000286711300023

    View details for PubMedID 21111698

  • V-gene amplification revisited - An optimised procedure for amplification of rearranged human antibody genes of different isotypes NEW BIOTECHNOLOGY Lim, T. S., Mollova, S., Rubelt, F., Sievert, V., Duebel, S., Lehrach, H., Konthur, Z. 2010; 27 (2): 108-117

    Abstract

    For studying human antibody variable (V)-gene usage in any group of individuals or for the generation of recombinant human antibody libraries for phage display, quality and yield of the amplified V-gene repertoire is of utmost importance. Key parameters affecting the amplification of full antibody repertoires are V-gene specific primer design, complementary DNA (cDNA) synthesis from total RNA extracts of peripheral blood mononuclear cells (PBMCs) and ultimately the polymerase chain reaction (PCR). In this work we analysed all these factors; we performed a detailed bioinformatic analysis of V-gene specific primers based on VBASE2 and evaluated the influence of different commercially available reverse transcriptases on cDNA synthesis and polymerases on PCR efficiency. The primers presented cover near to 100% of all functional and putatively functional V-genes in VBASE2 and the final protocol presents an optimised combination of commercial enzymes and reaction additives for cDNA synthesis and PCR conditions for V-gene amplification. Finally, applying this protocol in combination with different immunoglobulin (Ig) chain specific reverse primers we were able to amplify rearranged antibody genes of different isotypes under investigation.

    View details for DOI 10.1016/j.nbt.2010.01.001

    View details for Web of Science ID 000279133600006

    View details for PubMedID 20083243

Stanford Medicine Resources: