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1.
Sci Rep ; 9(1): 16468, 2019 11 11.
Article in English | MEDLINE | ID: mdl-31712621

ABSTRACT

All known methods for solid-phase synthesis of molecular arrays exploit positioning techniques to deposit monomers on a substrate preferably high densely. In this paper, stochastic patterning of molecule spots (250 000 spots monomers/cm2) via random allocation of the microbeads on a microstructured glass is presented. The size and shape of the microbeads and the microcavities are selected in such a way so that only one microbead can fit into the respective microcavity. Each microbead can be loaded with a certain type of molecule e.g. amino acids and is brought in the microcavities stochastically. Applying solvent vapor and heating the substrate, the molecules are released from the microbeads and coupled to the functionalized substrate. To differentiate between the microbeads carrying different molecules, quantum dot labels are preliminary introduced into the microbeads. Fluorescence imaging and subsequent data analysis enable decoding of the molecule deposition patterns. After the coupling step is completed, the microbeads are mechanically removed from the microwells. The composition of the monomer microbeads, their deposition and the conditions of the monomer extraction are studied. The stochastic monomer patterning may be used to design novel molecular arrays.


Subject(s)
Amino Acids/chemistry , Amino Acids/metabolism , Biosensing Techniques/methods , Fluorescence , Microarray Analysis/instrumentation , Microspheres , Quantum Dots , Solid-Phase Synthesis Techniques/methods , Humans
2.
Int J Mol Sci ; 20(13)2019 Jul 02.
Article in English | MEDLINE | ID: mdl-31269680

ABSTRACT

Much of the experimental data, especially in life sciences, is considered to be useless if it demonstrates a large standard deviation from the mean value. The Renaissance distribution, as presented in this study, allows one to extract true values from such statistical data with large noise. To obtain proof of the Renaissance distribution, high-throughput synthesis of deep substitutions for a target amino acid sequence was performed, and the known epitope was identified in assay with human serum antibodies. In addition, the Renaissance distribution was shown to approach the epitope affinity maturation by the deep alanine substitution. The Renaissance distribution may have an impact in the development of novel specific drugs.


Subject(s)
Alanine/genetics , Amino Acid Substitution , Peptides/genetics , Alanine/chemistry , Amino Acid Sequence , Epitopes/chemistry , Epitopes/genetics , Humans , Models, Molecular , Peptides/chemistry , Protein Array Analysis , Statistical Distributions , Stochastic Processes
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