Affinity capture of proteins from solution and their dissociation by contact printing
| Autor: | Hans Rudolf Bosshard, Emmanuel Delamarche, Hans Biebuyck, Peter Sonderegger, Dora Fitzli, Bruno Michel, André Bernard |
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| Přispěvatelé: | University of Zurich, Bernard, A |
| Rok vydání: | 2001 |
| Předmět: |
Time Factors
Biomedical Engineering Cell Culture Techniques 2204 Biomedical Engineering Bioengineering Chick Embryo Elastomer Transfection Applied Microbiology and Biotechnology Models Biological Dissociation (chemistry) chemistry.chemical_compound Affinity chromatography 10019 Department of Biochemistry Molecule 2402 Applied Microbiology and Biotechnology Animals Cells Cultured Neurons 1502 Bioengineering Dose-Response Relationship Drug Cell adhesion molecule Ligand Cell Adhesion Molecules Neuron-Glia Proteins Axons Kinetics chemistry Microscopy Fluorescence 1313 Molecular Medicine Immunoglobulin G COS Cells Biophysics 1305 Biotechnology 570 Life sciences biology Molecular Medicine Polystyrenes Polystyrene Contact print Biotechnology Protein Binding |
| Zdroj: | Nature biotechnology. 19(9) |
| ISSN: | 1087-0156 |
| Popis: | Biological experiments at the solid/liquid interface, in general, require surfaces with a thin layer of purified molecules, which often represent precious material. Here, we have devised a method to extract proteins with high selectivity from crude biological sample solutions and place them on a surface in a functional, arbitrary pattern. This method, called affinity-contact printing (alphaCP), uses a structured elastomer derivatized with ligands against the target molecules. After the target molecules have been captured, they are printed from the elastomer onto a variety of surfaces. The ligand remains on the stamp for reuse. In contrast with conventional affinity chromatography, here dissociation and release of captured molecules to the substrate are achieved mechanically. We demonstrate this technique by extracting the cell adhesion molecule neuron-glia cell adhesion molecule (NgCAM) from tissue homogenates and cell culture lysates and patterning affinity-purified NgCAM on polystyrene to stimulate the attachment of neuronal cells and guide axon outgrowth. |
| Databáze: | OpenAIRE |
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