Felix Löffler

High-Precision Combinatorial Deposition of Micro Particle Patterns on a Microelectronic Chip

The behavior of charged bio polymer micro particles when deposited onto a CMOS chip can be analytically modeled in form of the incompressible Navier-Stokes equation and the electrostatic Poisson equation, as we describe in this article. Based on these models, numerical simulations of depositions can be implemented in COMSOL that lead to improvements in the experimental setup, optimizing the size and charge distribution of the micro particles. Adapting the experiments according to the simulation results, we will show the powerful gain in deposition precision, which is essential for a contamination-free deposition and hence high quality combinatorial deposition.

Noncontact charge measurement of moving microparticles contacting dielectric surfaces

In this study examples for a noncontact procedure that allow the description of instant electric charging of moving microparticles that contact dielectric surfaces, for instance, of a flow hose are presented. The described principle is based on the measurement of induced currents in grounded metal wire probes, as moving particles pass close to the probe. The feasibility of the approach was tested with laser printer toner particles of a given size for different basic particle flow and charging conditions. An analytic description for the induced currents was developed and compared to observed effects in order to interpret the results qualitatively. The implementation of the presented procedure can be applied to transparent and nontransparent particle containers and flow lines of complex geometry which can be composed from the presented basic flow stream configurations.

Quality analysis of selective microparticle deposition on electrically programmable surfaces

Image processing and pattern analysis can evaluate the deposition quality of triboelectrically charged microparticles on charged surfaces. The image processing method presented in this paper aims at controlling the quality of peptide arrays generated by particle based solid phase Merrifield combinatorial peptide synthesis. Incorrectly deposited particles are detected before the amino acids therein are coupled to the growing peptide. The calibration of the image acquisition is performed in a supervised training step in which all parameters of the quality analyzing algorithm are learnt given one representative image. Then, the correct deposition pattern is determined by a linear support vector machine. Knowing the pattern, contaminated areas can be detected by comparing the pattern with the actual deposition. Taking into account the resolution of the image acquisition system and its magnification factor, the number and size of contaminating particles can be calculated out of the number of connected foreground pixels.

Peptide arrays with a chip.

Today, lithographic methods enable combinatorial synthesis of >50,000 oligonucleotides per cm(2), an advance that has revolutionized the whole field of genomics. A similar development is expected for the field of proteomics, provided that affordable, very high-density peptide arrays are available. However, peptide arrays lag behind oligonucleotide arrays. This is mainly due to the monomer-by-monomer repeated consecutive coupling of 20 different amino acids associated with lithography, which adds up to an excessive number of coupling cycles. A combinatorial synthesis based on electrically charged solid amino acid particles resolves this problem. A computer chip consecutively addresses the different charged particles to a solid support, where, when completed, the whole layer of solid amino acid particles is melted at once. This frees hitherto immobilized amino acids to couple all 20 different amino acids in one single coupling reaction to the support. The method should allow for the translation of entire genomes into a set of overlapping peptides to be used in proteome research.

Identification of a Tetanus Toxin Specific Epitope in Single Amino Acid Resolution

Vaccinations are among the most potent tools to fight infectious diseases. However, cross‐reactions are an ongoing problem and there is an urgent need to fully understand the mechanisms of the immune response. For the development of a methodological workflow, the linear epitopes in the immune response to the tetanus toxin is investigated in sera of 19 vaccinated Europeans applying epitope mapping with peptide arrays. The most prominent epitope, appearing in nine different sera (923IHLVNNESSEVIVHK937), is investigated in a substitution analysis to identify the amino acids that are crucial for the binding of the corresponding antibody species − the antibody fingerprint. The antibody fingerprints of different individuals are compared and found to be strongly conserved (929ExxEVIVxK937), which is astonishing considering the randomness of their development. Additionally, the corresponding antibody species is isolated from one serum with batch chromatography using the amino acid sequence of the identified epitope and the tetanus specificity of the isolated antibody is verified by ELISA. Studying antibody fingerprints with peptide arrays should be transferable to any kind of humoral immune response toward protein antigens. Furthermore, antibody fingerprints have shown to be highly disease‐specific and, therefore, can be employed as reliable biomarkers enabling the study of cross‐reacting antigens.

Kombinatorische Chemie im hochdichten Arrayformat

We synthesize peptide arrays by using laser pulses to transfer amino acids onto a synthesis slide that are embedded in a polymer. After patterning these razor-thin spots, the coupling starts by heating the polymer that now serves as a solvent to couple monomers to the surface. The main advantages of this nanolayer solid phase chemistry are the high degree of automation, the very high spot density, and the in situ activation of monomers due to mixing different reagents in the stacked nanolayers.

Image Processing Quality Analysis for Particle Based Peptide Array Production on a Microchip

Highly complex microarray systems based on combinatorial synthesis techniques are in wide-spread use in biological, medical and pharmaceutical research Chee et al. (1996); Cretich et al. (2006); Debouck & Goodfellow (1999). Two prominent examples are micro arrays for the artificial synthesis of arbitrary DNA sequences out of nucleic acids Heller (2002) and peptide synthesis out of amino acids Beyer et al. (2007); Templin et al. (2003). In the case of DNA arrays, these experiments mostly focus on gene identification or gene expression profiling to determine the effects of single genes on cellular evolution. Peptide arrays aim at understanding interactions of peptides with other molecules. As sequences in proteins, peptides are involved in the regularisation of biological activity.