Microstructure-Assisted Particle Pattern Generation for High Density Peptide Arrays

We developed particle patterning processes on microstructured surfaces for the combinatorial synthesis of high density peptide arrays. We worked with polymer microparticles with embedded amino acids derivatives and solid substrates with cylindrical microcavities. Using laser-based method, we demonstrated combinatorial particle patterning with 1 million spots/cm². With the so-called stochastic approach, we synthesized dipeptide array with a density of 25 million peptide spots/cm²

The Functional Activity of Neutrophils in Athletes

Introduction- Polymorphonuclear neutrophils (PMNs) take direct part in reaction of whole body on exercise. The aim of present study was to investigate the functional activity of PMNs in athletes. Methods - Quantity of PMNs, their phagocytic and oxidative activity and capacity to produce interleukin-6 (IL-6) and IL-8 in vitro in the presence or absence of stimuli (LPS and zymosan A) was studied in 3 groups of athletes parted on the basis of the clinical and anamnestic data: group 1 (n=27) - athletes without the clinical diagnosis (healthy); group 2 (n=11) with persistent infections in the anamnesis (a herpes simplex and influenza virus, a genyantritis, a tonsillitis, etc.); group 3 (n=8) with sings of any allergic responses in the anamnesis. The healthy untrained volunteers served as the controls (n=109). Results– Athletes with allergy had a lower relative quantity of PMNs than in controls (р=0.019) and decreased absolute quantity of PMNs in comparison with healthy athletes (р=0.045). PMNs in athletes had decreased phagocytic activity. However phagocytic capacity of PMNs in athletes from groups 2 and 3 was suppressed significantly (1.6- and 2.1-fold, respectively, р=0.001, р=0.0001) in comparison with controls. Athletes with allergy had also a significantly lower phagocytic activity of PMNs than healthy athletes. PMNs in athletes had raised spontaneous respiratory burst (quantity of formazan-positive cells in NBT-test, FPC) than in controls (4-, 10- and 2.5-fold for groups 1-3, respectively, р=0.0001, р=0.0001, р=0.014). Healthy athletes had elevated quantity of FPC after stimulation of PMNs by zymosan (р=0.05). At the same time stimulated oxidative activity of PMNs in athletes from groups 2 and 3 was suppressed. PMNs in healthy athletes possessed the raised ability to produce of IL-6 and IL-8 spontaneously and after stimulation in comparison with controls (all р Conclusion - Our data show that regular exercise influence on key effector function of PMNs and its capacity to produce the cytokines. We also observed more significant changes in the functional activity of PMNs in athletes with persistent infections and allergy in the anamnesis. Possibly these changes are linked with a higher incidence of infection and allergic responses in some athletes

Selective Functionalization of Microstructured Surfaces by Laser-Assisted Particle Transfer

Microcavity arrays represent millions of different reaction compartments to screen for e.g. molecular interactions, exogenous factors for cells or enzymatic activity. We present a novel method to selectively synthesize different compounds in arrays of microcavities with up to 1,000,000 cavities per cm2. In our approach, polymer microparticles with embedded pre-activated monomers are selectively transferred into microcavities with laser radiation. After particle patterning, heating of the particle matrix simultaneously leads to diffusion and coupling of the monomers inside each microcavity separately. This method exhibits flexibility, not only in the choice of compounds, but also in the choice of particle matrix material, which determines the chemical reaction environment. The laser-assisted selective functionalization of microcavities can be easily combined with the intensively growing number of laser applications for patterning of molecules and cells, which is useful for the development of novel biological assays

High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in solid matrix material

Laser writing is used to structure surfaces in many different ways in materials and life sciences. However, combinatorial patterning applications are still limited. Here we present a method for cost-efficient combinatorial synthesis of very-high-density peptide arrays with natural and synthetic monomers. A laser automatically transfers nanometre-thin solid material spots from different donor slides to an acceptor. Each donor bears a thin polymer film, embedding one type of monomer. Coupling occurs in a separate heating step, where the matrix becomes viscous and building blocks diffuse and couple to the acceptor surface. Furthermore, we can consecutively deposit two material layers of activation reagents and amino acids. Subsequent heat-induced mixing facilitates an in situ activation and coupling of the monomers. This allows us to incorporate building blocks with click chemistry compatibility or a large variety of commercially available non-activated, for example, posttranslationally modified building blocks into the array’s peptides with >17,000 spots per cm²

Particle-Based Microarrays of Oligonucleotides and Oligopeptides

In this review, we describe different methods of microarray fabrication based on the use of micro-particles/-beads and point out future tendencies in the development of particle-based arrays. First, we consider oligonucleotide bead arrays, where each bead is a carrier of one specific sequence of oligonucleotides. This bead-based array approach, appearing in the late 1990s, enabled high-throughput oligonucleotide analysis and had a large impact on genome research. Furthermore, we consider particle-based peptide array fabrication using combinatorial chemistry. In this approach, particles can directly participate in both the synthesis and the transfer of synthesized combinatorial molecules to a substrate. Subsequently, we describe in more detail the synthesis of peptide arrays with amino acid polymer particles, which imbed the amino acids inside their polymer matrix. By heating these particles, the polymer matrix is transformed into a highly viscous gel, and thereby, imbedded monomers are allowed to participate in the coupling reaction. Finally, we focus on combinatorial laser fusing of particles for the synthesis of high-density peptide arrays. This method combines the advantages of particles and combinatorial lithographic approaches

High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in solid matrix material

International audienceLaser writing is used to structure surfaces in many different ways in materials and life sciences. However, combinatorial patterning applications are still limited. Here we present a method for cost-efficient combinatorial synthesis of very-high-density peptide arrays with natural and synthetic monomers. A laser automatically transfers nanometre-thin solid material spots from different donor slides to an acceptor. Each donor bears a thin polymer film, embedding one type of monomer. Coupling occurs in a separate heating step, where the matrix becomes viscous and building blocks diffuse and couple to the acceptor surface. Furthermore, we can consecutively deposit two material layers of activation reagents and amino acids. Subsequent heat-induced mixing facilitates an in situ activation and coupling of the monomers. This allows us to incorporate building blocks with click chemistry compatibility or a large variety of commercially available non-activated, for example, posttranslationally modified building blocks into the array's peptides with 417,000 spots per cm(2)