Structure-based design of prefusion-stabilized SARS-CoV-2 spikes.
Hsieh Ching-Lin, Goldsmith Jory A, Schaub Jeffrey M, DiVenere Andrea M, Kuo Hung-Che, Javanmardi Kamyab, Le Kevin C, Wrapp Daniel, Lee Alison G, Liu Yutong, Chou Chia-Wei, Byrne Patrick O, Hjorth Christy K, Johnson Nicole V, Ludes-Meyers John, Nguyen Annalee W, Park Juyeon, Wang Nianshuang, Amengor Dzifa, Lavinder Jason J, Ippolito Gregory C, Maynard Jennifer A, Finkelstein Ilya J, McLellan Jason S
Science (New York, N.Y.) · 2020 · PMID 32703906
The coronavirus disease 2019 (COVID-19) pandemic has led to accelerated efforts to develop therapeutics and vaccines. A key target of these efforts is the spike (S) protein, which is metastable and difficult to produce recombinantly. We characterized 100 structure-guided spike designs and identified 26 individual substitutions that increased protein yields and stability.
Testing combinations of beneficial substitutions resulted in the identification of HexaPro, a variant with six beneficial proline substitutions exhibiting higher expression than its parental construct (by a factor of 10) as well as the ability to withstand heat stress, storage at room temperature, and three freeze-thaw cycles. A cryo-electron microscopy structure of HexaPro at a resolution of 3.2 angstroms confirmed that it retains the prefusion spike conformation. High-yield production of a stabilized prefusion spike protein will accelerate the development of vaccines and serological diagnostics for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).