Single-Chain Antibody Library
A PNNL scientist uses a BD Biosciences high-performance flow cytometer, the FACSAria, for human antibody studies.
Researchers at Pacific Northwest National Laboratory (PNNL) have constructed a nonimmune library consisting of 109 human antibody scFv fragments, which have been cloned and expressed on the surface of yeast. Nanomolar-affinity scFvs are routinely obtained by magnetic bead screening and flow cytometric sorting. The yeast library can be amplified 1010 fold without measurable loss of clonal diversity. This allows for indefinite expansion of the library. All scFv clones can be assessed directly on the yeast cell surface by immunofluorescent labeling and flow cytometry, obviating separate subcloning, expression, and purification steps. The ability to use multiplex library screening demonstrates the utility of this approach for high-throughput antibody isolation for proteomic applications.
The yeast library may be used for research projects or teaching performed for U.S. Government purposes only. If you would like to request an aliquot of the single-chain antibody library for your research, please print and fill out the Materials Transfer Agreement (MTA) [PDF, 19KB] and send it to:
Pacific Northwest National Laboratory
902 Battelle Boulevard
P.O. Box 999, MSIN K1-53
Richland, WA 99352
Material will only be sent to those who provide a Fed Ex number on the MTA. PNNL will review requests and reserves the right to deny any requests that do not meet the criteria listed on the MTA. For questions, call Bob at 509/375-6518.
- Yeast Display scFV Antibody Library User's Manual [PDF, 557KB]
- Genetic sequence of pPNL6 and pPNL9 - to view these files you must have Vector NTI software installed on your computer.
- Frequently Asked Questions
- MIT collaborator, Dane Wittrup Lab
- Miltenyi Biotec's display technology
- Library aliquots received before 2007 may be contaminated with a competing yeast strain. Read about the potential problem: Candida Contamination in the scFv Yeast Display Library and Methods for it's Control [PDF, 235KB].
- Protocol for making targeted immune yeast display libraries.
For technical questions, fill out the information request form.