Coronaviridae spike glycoproteins mediate viral entry and fusion to host cells through binding to host receptors (i.e., ACE2, DPP4) and are key components in determining viral host range, making them targets for antiviral research. Here, we describe the expression, purification, and characterization of recombinant spike proteins to aid in protein characterization and analysis. These protocols were used for the production of spike glycoproteins from civet, pangolin, and bat coronaviruses, as well as high-resolution cryo-electron microscopy (cryo-EM) structural analysis of bat and civet host coronavirus spike glycoproteins (Hills et al., 2024). © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Expression and purification of SARS-CoV spike protein from ExpiCHO cells Basic Protocol 2: Preparation of SARS-CoV spike protein for visualization by negative-stain transmission electron microscopy and cryo-electron microscopy.
Download full-text PDF |
Source |
---|---|
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11890022 | PMC |
http://dx.doi.org/10.1002/cpz1.70115 | DOI Listing |
Front Immunol
March 2025
Abterra Biosciences, Inc., San Diego, CA, United States.
The rapid spread of SARS-CoV-2 and its continuing impact on human health has prompted the need for effective and rapid development of monoclonal antibody therapeutics. In this study, we investigate polyclonal antibodies in serum and B cells from the whole blood of three donors with SARS-CoV-2 immunity to find high-affinity anti-SARS-CoV-2 antibodies to escape variants. Serum IgG antibodies were selected by their affinity to the receptor-binding domain (RBD) and non-RBD sites on the spike protein of Omicron subvariant B.
View Article and Find Full Text PDFFront Immunol
March 2025
R&D Laboratory, Diagnosticum Zrt, Budapest, Hungary.
Antigen specific humoral immunity can be characterized by the analysis of serum antibodies. While serological assays for the measurement of specific antibody levels are available, these are not quantitative in the biochemical sense. Yet, understanding humoral immune responses quantitatively on the systemic level would need a universal, complete, quantitative, comparable measurement method of antigen specific serum antibodies of selected immunoglobulin classes.
View Article and Find Full Text PDFFront Immunol
March 2025
Department of Vaccine Research and Development, Applied Biomedical Science Institute, San Diego, CA, United States.
The SARS-CoV-2 pandemic, while subsiding, continues to plague the world as new variants emerge. Millions have died, and millions more battle with the debilitating symptoms of a clinical entity known as long Covid. The biggest challenge remains combating an ever-changing variant landscape that threatens immune evasion from vaccine and prior infection-generated immunity.
View Article and Find Full Text PDFArch Microbiol
March 2025
Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, No. 22, Jinjing Road, Xiqing District, Tianjin, 300392, People's Republic of China.
Avian infectious bronchitis (IB) is one of the major respiratory diseases in poultry. At present, attenuated vaccines are the main commercial vaccines, but they have many defects. We aimed to construct a novel multi-epitope DNA vaccine based on avian infectious bronchitis virus (IBV) S1 and N proteins for the prevention of IBV infection.
View Article and Find Full Text PDFJ Korean Med Sci
March 2025
Emerging Infectious Diseases Research Institute, Chungnam National University Hospital, Daejeon, Korea.
Background: We evaluated severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2)-specific humoral and cellular responses for up to 6 months after the 3rd dose of ancestral coronavirus disease 2019 (COVID-19) vaccination in people living with HIV (PLWH) and healthy controls (HCs) who were not infected with COVID-19.
Methods: Anti-spike receptor-binding domain IgG (anti-RBD IgG) concentrations using chemiluminescence immunoassay and neutralizing antibodies using focus reduction neutralization test (FRNT) were assessed at 1 week after each dose of vaccination, and 3 and 6 months after the 3rd dose in 62 PLWH and 25 HCs. T-cell responses using intracellular cytokine stain were evaluated at 1 week before, and 1 week and 6 months after the 3rd dose.
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!