C. Hsieh, J. Goldsmith, Jeffrey M. Schaub, Andrea M DiVenere, Hung-Che Kuo, Kamyab Javanmardi, Kevin C. Le, D. Wrapp, A. G. Lee, Yutong Liu, Chia-Wei Chou, P. O. Byrne, Christy K. Hjorth, Nicole V. Johnson, J. Ludes-Meyers, Annalee W. Nguyen, Juyeon Park, Nianshuang Wang, Dzifa Amengor, J. Maynard, I. J. Finkelstein, J. McLellan
bioRxiv : the preprint server for biology, 2020
The COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2 has led to accelerated efforts to develop therapeutics, diagnostics, and vaccines to mitigate this public health emergency. A key target of these efforts is the spike (S) protein, a large trimeric class I fusion protein that is metastable and difficult to produce recombinantly in large quantities. Here, we designed and expressed over 100 structure-guided spike variants based upon a previously determined cryo-EM structure of the prefusion SARS-CoV-2 spike. Biochemical, biophysical and structural characterization of these variants identified numerous individual substitutions that increased protein yields and stability. The best variant, HexaPro, has six beneficial proline substitutions leading to ∼10-fold higher expression than its parental construct and is able to withstand heat stress, storage at room temperature, and multiple freeze-thaws. A 3.2 Å-resolution cryo-EM structure of HexaPro 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 SARS-CoV-2.
Cited by 154 publications.
Field of study: Medicine