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Expanding and improving nanobody repertoires using a yeast display method: Targeting SARS-CoV-2.
Frederick R Cross1, Peter C Fridy2, Natalia E Ketaren2
1Laboratory of Cell Cycle Genetics, The Rockefeller University, New York, New York, USA.
The Journal of Biological Chemistry
|January 31, 2023
Summary
Researchers optimized yeast display to discover potent nanobodies against SARS-CoV-2 variants. This method efficiently identifies diverse nanobodies with high neutralization activity, offering a promising therapeutic strategy against COVID-19.
Area of Science:
- Immunology
- Virology
- Biotechnology
Background:
- COVID-19, caused by SARS-CoV-2, poses a global health challenge with evolving variants.
- Nanobodies, derived from camelids, show potential as therapeutics due to their small size and high affinity.
- Developing effective nanobodies against SARS-CoV-2 variants is crucial for ongoing therapeutic strategies.
Purpose of the Study:
- To optimize a yeast display method for identifying and characterizing a broad range of anti-SARS-CoV-2 Spike nanobodies.
- To compare the efficiency and scope of the optimized yeast display method with a mass spectrometry-based approach.
- To explore the potential of DNA shuffling in enhancing nanobody function.
Main Methods:
- Optimized a yeast display platform using B-cell cDNA from immunized animals to generate VHH sequences.
- Leveraged a previously established mass spectrometry-based method for comparison.
- Applied DNA shuffling to complementarity-determining regions (CDRs) of nanobodies.
- Utilized the yeast display for massively parallel epitope binning.
Main Results:
- The optimized yeast display successfully identified numerous nanobody sequences, including many not found by the mass spectrometry method.
- Newly identified nanobodies demonstrated high affinity and potent neutralization activity against diverse SARS-CoV-2 variants.
- DNA shuffling indicated a modular function of CDRs in antiviral nanobodies.
- Yeast display proved comparable in efficiency and specificity to mass spectrometry but required less specialized infrastructure.
Conclusions:
- The optimized yeast display method provides a powerful, accessible platform for comprehensively exploring nanobody repertoires against SARS-CoV-2.
- This approach significantly increases the likelihood of discovering broadly specific nanobodies and synergistic combinations for COVID-19 therapy.
- Yeast display and mass spectrometry are complementary techniques for deep paratope exploration.
