Panning of a phage VH library using nitrocellulose membranes: application to selection of a human VH library.

摘要: We have established a method for selecting binding phages from phage immunoglobulin heavy chain variable region (VH) library by panning with nitrocellulose membranes (membrane panning). To evaluate the concentrating ability of membrane phages, VH containing clones that bind to hen egg white lysozyme (HEL) was used against HEL. The efficiency our as high magnetic beads. In addition, we performed target proteins and isolated phages. human genes these were cloned expressed VH-bacterial alkaline phosphatase (PhoA) conjugates (VH-PhoA) in Escherichia coli. dose-dependent VH-PhoA confirmed dot blotting. When applied disease-associated antibodies, methods will likely benefit clinical research. techniques may be applicable systematic analysis proteome studies.