Affinity capture of proteins from solution and their dissociation by contact printing.
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.
Bernard
A
A
IBM Research, Zurich Research Laboratory, Säumerstr. 4, CH-8803 Rüschlikon, Switzerland. a.bernard@lifebits.de
Fitzli
D
D
Sonderegger
P
P
Delamarche
E
E
Michel
B
B
Bosshard
H R
HR
Biebuyck
H
H
D000818:Animals; D001369:Axons / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D019556:COS Cells; D019062:Cell Adhesion Molecules, Neuron-Glia / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D007074:Immunoglobulin G / Q000378:metabolism; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism; D011137:Polystyrenes / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification; D013997:Time Factors; D014162:Transfection
D019062:Cell Adhesion Molecules, Neuron-Glia; D007074:Immunoglobulin G; D011137:Polystyrenes; D011506:Proteins