Flow chart of the capture-lift screening procedure. Nitrocellulose filters are incubated with a capture reagent (1), then the remaining nonspecific binding sites on the filter are blocked (2). A bacterial lawn is infected with the Ab-phage library (3) and is overlaid with the pretreated nitrocellulose filter for 12–14 h (4). The filter is removed and probed with labeled Ag(s) and the appropriate detection reagent (5). In the example in this figure, three distinct Fabs are reactive with target A, but two are crossreactive with targets B and C. The developed filter is aligned with the agar plate to isolate the clone(s) of interest specifically reactive with target A (6).
Briefly, the nitrocellulose filters are coated with an Ab capture
reagent, such as a polyclonal anti-κ-chain Ab (for human Fabs) and
the remaining protein-binding sites are blocked prior to performing
the plaque lift ( Fig.
Viewing this image requires a subscription. If you are a subscriber, please log in.
This image is from the chapter titled "Screening of Phage-Expressed Antibody Libraries by Capture Lift"
(from Antibody Phage Display), which is copyrighted by Humana Press Inc. For more information on the
copyright for this image, please refer to the full image caption and to the
The image is being made available for non-commercial purposes for subscribers to SpringerImages. For more information on what you are allowed to do with this image, please see our copyright policy.
To request permissions to use any copyrighted material, please visit the source document.
Report a copyright concern regarding this image.
Log in or register to save your favorite images and download them as high-quality PowerPoint or PDF files.
Log in or register to save your search criteria.
© Springer, part of Springer Science+Business Media.
Remote Address: 220.127.116.11 Server: 20