Title
Production of novel SARS-CoV-2 Spike truncations in Chinese hamster ovary cells leads to high expression and binding to antibodies
Author
Shiaki A. Minami, Seongwon Jung, Yihan Huang, Bradley S. Harris, Matthew W. Kenaston, Roland Faller, Somen Nandi, Karen A. McDonald, Priya S. Shah
Year
2022
Journal
Biotechnology Journal
Abstract
SARS-CoV-2 Spike is a key protein that mediates viral entry into cells and elicits antibody responses. Its importance in infection, diagnostics, and vaccinations has created a large demand for purified Spike for clinical and research applications. Spike is difficult to express, prompting modifications to the protein and expression platforms to improve yields. Alternatively, the Spike receptor-binding domain (RBD) is commonly expressed with higher titers, though it has lower sensitivity in serological assays. Here, we improve transient Spike expression in Chinese hamster ovary (CHO) cells. We demonstrate that Spike titers increase significantly over the expression period, maximizing at 14 mg L−1 on day 7. In comparison, RBD titers peak at 54 mg L−1 on day 3. Next, we develop eight Spike truncations (T1–T8) in pursuit of truncation with high expression and antibody binding. The truncations T1 and T4 express at 130 and 73 mg L−1, respectively, which are higher than our RBD titers. Purified proteins were evaluated for binding to antibodies raised against full-length Spike. T1 has similar sensitivity as Spike against a monoclonal antibody and even outperforms Spike for a polyclonal antibody. These results suggest that T1 is a promising Spike alternative for use in various applications.
Instrument
J-715
Keywords
SARS-CoV-2, spike protein, RBD, protein