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Phage Display Plasmids with Improved Expression Properties for Human and Chimeric Nonhuman/Human Fab Libraries
| | Inventor: Christoph Rader (NCI) | Record Date: 11/17/2009 | | Last Updated: 11/17/2009 |
Abstract: | | The Fab molecule was the first generated antibody fragment and still dominates basic research and clinical applications. New phage display vectors were designed to generate and select Fab libraries with human constant domains. These vectors facilitate bacterial expression of human, humanized, and chimeric nonhuman/human Fab antibody fragments. They differ from currently available pComb3H and pComb3X phage display vectors by assembling human and chimeric nonhuman/human Fab libraries in two rather than three PCR steps. As a result, these novel constructs retain the initial variable light and heavy chain sequences and improve the resulting Fab library’s complexity in terms of number, diversity, and affinity. These constructs were developed with and without a His tag and yield approximately 100 µg to 2 mg of protein, which can be used for evaluation and characterization of Fab binding properties such as affinity and specificity. Notably, the His tag provides a handle to easily purify Fab. Advantages: -- Improved Fab library with complexity and number, diversity, and affinity; -- His tag construct allows for simplified purification assays |
Applications: | | Generation of human, humanized, and chimeric nonhuman/human Fab antibody fragments Research tool to characterize Fab antibody fragments |
Patent:
HHS Reference No. E-008-2010/0 – Research Tool. Patent protection is not being pursued for this technology. |
License:
Available for licensing. |
Address:
National Institutes of Health
Office of Technology Transfer
6011 Executive Boulevard, Suite 325
Rockville, MD 20852
USA
Phone: 301-435-4633
Fax: 301-402-0220 | |
E-mail:
wongje@mail.nih.gov
URL:
http://www.ott.nih.gov
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