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Revision as of 18:18, 25 January 2021
Contents
Module 3
Lecturer: Dr. Leslie McClain
Instructors: Dr. Noreen Lyell and Dr. Leslie McClain, and Dr. Becky Meyer
Research assistant: Sarah Cowles
TAs: Jeff Hsaio and Malek Kabani
Engineering Antibodies Using Yeast Display
An antibody is a soluble or membrane bound protein, produced by immune cells in blood. In our bodies the purpose of an antibody is to recognize foreign substances, called antigens, and mark them for removal by cells that compromise our immune system. If one has a properly functioning immune system you've all been making new and improved antibodies your entire life.
Antibodies are also a critical reagent in medicine, research and diagnostics. Scientists have found ways to induce and purify antibodies from animals, and to also engineer cells to produce specific antibodies. In Mod1 we'll use a method developed in the Wittrup lab to screen a yeast library of antibodies, against a particular antigen, using Fluorescence Assisted Cell Sorting, and try to identify a single antibody that shows improved antigen binding in a quantitative binding assay.
This module was developed thanks to the invaluable help and support of Wittrup lab PhD student, Sarah Cowles, and the generous contribution of reagents from Prof. Wittrup.
Research goal:
Lab links: day by day
M1D1: [[]]
M1D2: [[]]
M1D3: [[]]
M1D4: [[]]
M1D5: [[]]
Assignments
References
- Isolating and engineering human antibodies using yeast surface display. Nature Protocols. 1, p755–768(2006).
- Yeast Surface Display for Antibody Isolation: Library Construction, Library Screening, and Affinity Maturation. Methods in Molecular Biology . 1131:151-81(2014).
- Applications of Yeast Surface Display for Protein Engineering. Methods in Molecular Biology (2015) 1319:155-75.
- Examples of Using Yeast Surface Display for Non-antibody Proteins:
- Stability and CDR Composition Biases Enrich Binder Functionality Landscapes. J. Mol. Biol. (2010) 401, 84–96
- Beyond Epitope Binning: Directed in Vitro Selection of Complementary Pairs of Binding Proteins. ACS Comb. Sci. (2020) 22, 49−60.
- Protocol for expressing full antibodies in mammalian cells:
- Transient expression of human antibodies in mammalian cells. Nature Protocols (2018) 13(10), 99-117