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Protein Engineering by Yeast Surface Display
Published on: November 29, 2024
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Precise and efficient antibody epitope determination through library design, yeast display and next-generation
Thomas Van Blarcom1, Andrea Rossi1, Davide Foletti1
1Rinat, Pfizer Inc., 230 East Grand Avenue, South San Francisco, CA 94080, USA.
Journal of Molecular Biology
|October 7, 2014
Summary
We developed a rapid, precise method to map antibody binding sites (epitopes) using yeast surface display and DNA sequencing. This technique helps predict antibody effectiveness against bacterial toxins like Staphylococcus aureus alpha toxin.
Area of Science:
- Biochemistry
- Immunology
- Protein Engineering
Background:
- Therapeutic antibodies are a rapidly growing class of drugs due to their high specificity and affinity.
- Precisely identifying antibody epitopes is crucial for understanding their mechanism of action and differentiating similar antibodies.
Purpose of the Study:
- To describe a novel method for precise and efficient parallel epitope mapping of multiple antibodies.
- To demonstrate the method's utility in characterizing antibodies targeting Staphylococcus aureus alpha toxin.
Main Methods:
- Combination of rational library design, quantitative yeast surface display, and next-generation DNA sequencing.
- Epitope mapping of antibodies against Staphylococcus aureus alpha toxin.
- Validation using co-crystal structure and antibody affinity variants.
Main Results:
- Accurate and efficient parallel epitope mapping was achieved.
- Quantitative insights into critical epitope residues and antibody-antigen interactions were obtained.
- Relative antibody affinities were estimated, predicting neutralizing potency against alpha toxin variants.
Conclusions:
- The developed method provides precise epitope information and predicts antibody efficacy.
- This approach aids in selecting optimal antibody candidates for therapeutic development.
- The method is applicable to antibodies targeting various antigens, including bacterial toxins.
