Aptares AG

Mittenwalde, Germany

Aptares AG

Mittenwalde, Germany
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Wallukat G.,Max Delbruck Centrum fur Molekulare Medizin | Wallukat G.,Aptares AG | Haberland A.,Charité - Medical University of Berlin | Berg S.,Institute For Diabetes Gerhardt Katsch | And 9 more authors.
Circulation Journal | Year: 2012

Background: Application of immunoapheresis to eliminate pathogenic autoantibodies targeting the second extracellular loop of the β1-receptor (β1-AABs) is currently investigated in patients with cardiomyopathy. Aptamers (single short DNA or RNA strands) are a new class of molecules that bind to a specific target molecule. This property qualifies aptamers for potential use in the apheresis technique. We recently identified an aptamer that specifically binds to β1-AABs, so in the present study we tested whether this aptamer could be used as a binder to prepare an apheresis column suitable for clearing β1-AABs from rat's blood. Methods and Results: An apheresis column was designed containing the β1-AAB-targeting-aptamer coupled to sepharose. As tested in vitro, this column (1) binds β1-AABs highly specifically without marked interference with common IgGs, (2) has a capacity for clearing of approximately 1L of β1-AAB-positive serum and (3) can be completely regenerated for subsequent use. Using the column for extracorporeal apheresis of spontaneously hypertensive rats (SHR) positive for both β1-AABs and muscarinic 2-receptor autoantibodies (M2-AABs), only β1-AABs were removed. In a follow-up of 9 weeks, recurrence of β1-AABs in the blood of SHR could not be detected. Conclusions: For the first time, a newly designed apheresis column with a β1-AAB specific aptamer as a binder was successfully used to eliminate β1-AABs from SHR blood.

In this project, we aim to develop nanotechnology-based systems for detection of pathogenic bacteria in several aqueous media mainly in environmental water based on suspension arrays together with a portable custom-designed detection system. The six objectives (six work packages) are as follows: (i) Nanostructured sensors surfaces, in which electron-beam, photo-, soft and dip-pen lithographies will be applied in the fabrication of functionalized low-cost (disposable) SERS substrates, specially designed for simultaneus Raman and Fluorescence enhancement; (ii) nanosorbents and their mixtures as suspension arrays against the target bacteria for capturing units in which firstly magnetically loaded nanoparticles will be produced and labelled/barcoaded with fluorosence dies and/or quantum dots and then bacteriophages, monoclonal antibodies, and/or aptamers will be immobilized onto these nanoparticles as bioligands to recognize and capture the target bacteria specifically; their mixtures will form the suspension arrays; (iii) target bacteria and bacteriophages; the target bacteria (as water pollutants), are Escherichia coli, Enterococcus species and Bacteroides ovatus; bacteriophages which will recognize these bacteria will be selected/produced as bioligands; (iv) aptamers; they will be selected/modified/produced; (v) sensors/array systems; a portable Raman Spectrometer/Fluorescence Detector System will be designed/produced that will be used together with the nanostructured sensors platforms, and finally (vi) validation; the materials and systems developed will be validated. This is a multidisciplinary and technological project and therefore brings together experts from different disciplines both from academia and industry. Basicly, the knowledge that will be developed/ accumulated at lab scale during the project will be transfered to the industrial partners for prototype productions, and then these will bring back to the academical institutions for validation tests.

Haberland A.,Charité - Medical University of Berlin | Wallukat G.,Max Delbruck Centrum Berlin | Wallukat G.,Aptares AG | Dahmen C.,Aptares AG | And 2 more authors.
Circulation Research | Year: 2011

Rationale Autoantibodies directed against the beta1-adrenoceptor (beta1-AABs) have been proposed to drive the pathogenesis of idiopathic dilated cardiomyoparthy (DCM), Chagas cardiomyopathy, and peripartum cardiomyopathy. For disease treatment, aptamers that bind and neutralize beta1-AABs could be significant. Objective: We determined whether oligonucleotide-aptamers, selected to target human beta1-AABs directed against the second extracellular loop of the beta1-AAB, can neutralize these AABs and modulate their function in vitro. Methods and Results: Using Monolex technology, we identified an ssDNA aptamer that targets human beta1-AABs. The neutralization potential of this aptamer against beta1-AABs isolated from patients with DCM, Chagas cardiomyopathy, and peripartum cardiomyopathy was analyzed using cultured neonatal rat cardiomyocytes by monitoring beta1-AAB induced cell toxicity and chronotropic cell responses. Aptamer addition reduced beta1-AAB induced cell toxicity and neutralized chonotropic beta1-AAB function in a dose-dependent manner. In the presence of aptamer neutralized beta1-AABs, cells remained fully responsive to agonists and antagonists, such as isoprenaline and bisoprolol. Both aptamer pretreated with a complementary (antisense) aptamer and a control scrambled-sequence aptamer were ineffective at beta1-AAB neutralization. Beta1-AABs directed against the first extracellular loop of the beta1-receptor and AABs directed against other G-protein coupled receptors were not affected by the selected aptamer. Conclusions: A specific aptamer that can neutralize cardiomyopathy associated human beta1-AABs in vitro has been identified and characterized, providing a framework for future in vivo testing of this treatment option in animal experiments. © 2011 American Heart Association. All rights reserved.

Negri P.,University of Georgia | Chen G.,University of Georgia | Kage A.,AptaRes AG | Nitsche A.,Robert Koch Institute | And 3 more authors.
Analytical Chemistry | Year: 2012

We have demonstrated label-free optical detection of viral nucleoprotein binding to a polyvalent anti-influenza aptamer by monitoring the surface-enhanced Raman (SERS) spectra of the aptamer-nucleoprotein complex. The SERS spectra demonstrated that selective binding of the aptamer-nucleoprotein complex could be differentiated from that of the aptamer alone based solely on the direct spectral signature for the aptamer-nucleoprotein complex. Multivariate statistical methods, including principal components analysis, hierarchical clustering, and partial least squares, were used to confirm statistically significant differences between the spectra of the aptamer-nucleoprotein complex and the spectra of the unbound aptamer. Two separate negative controls were used to evaluate the specificity of binding of the viral nucleoproteins to this aptamer. In both cases, no spectral changes were observed that showed protein binding to the control surfaces, indicating a high degree of specificity for the binding of influenza viral nucleoproteins only to the influenza-specific aptamer. Statistical analysis of the spectra supports this interpretation. AFM images demonstrate morphological changes consistent with formation of the influenza aptamer-nucleoprotein complex. These results provide the first evidence for the use of aptamer-modified SERS substrates as diagnostic tools for influenza virus detection in a complex biological matrix. © 2012 American Chemical Society.

Negri P.,University of Georgia | Kage A.,AptaRes AG | Nitsche A.,Robert Koch Institute | Naumann D.,Robert Koch Institute | Dluhy R.A.,University of Georgia
Chemical Communications | Year: 2011

A highly sensitive surface-enhanced Raman (SERS)-based method for detection of influenza viral nucleoproteins is described. The intrinsic SERS spectrum of the aptamer-nucleoprotein complex provides direct evidence of binding between a polyvalent anti-influenza aptamer and the nucleoproteins of three influenza strains. © The Royal Society of Chemistry 2011.

The present invention is directed to aptamers that are characterized in that it inhibits the agonistic effect of an antibody specific for the 1st or 2nd extracellular loop of human beta-1-adrenergic receptor in a rat cardiomyocyte beating frequency assay with an IC50 of 100 nM or less.

The invention relates to a method for selecting nucleic acids that bond with high affinity to a target molecule from a mixture of nucleic acids, comprising: a) loading a column with the target molecules, b) feeding the nucleic acids into a first end of the column, to create a defined volumetric flow of medium through the column, c) immobilizing the nucleic acids to the target molecule wherein an affinity of the nucleic acids to the target molecule decreases as the distance from the first end of the column increases, d) stopping the volumetric flow after a defined period of time, e) cutting the column into segments, and allocating a routing co-ordinate to each segment, and f) identifying and collecting nucleic acids that bond with a high affinity to the target molecule by desorbing the immobilized nucleic acids from at least one segment.

The invention relates to a method of selection, by two-dimensional separation, of nucleic acids that bind to a target molecule with high affinity from a mixture of nucleic acids, including the following steps: a) subjecting the mixture of nucleic acids to a physico-chemical separation step, thereby obtaining a set of mixed fractions containing the nucleic acids, a run parameter window being associated with every mixed fraction containing the nucleic acids, b) contacting a mixed fraction containing the nucleic acids with the target molecule, thereby obtaining a binding mixture containing nucleic acid/target molecule complexes, c) subjecting the binding mixture from step b) to the same physico-chemical separation step as in step a), thereby selecting nucleic acid/target molecule complexes whose run parameters are outside of the run parameter window.

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