Recombinant Severe acute respiratory syndrome coronavirus 2 Nucleoprotein (N) (Q384H) is expressed in E. coli, covering the full-length sequence from 1-419 amino acids with a Q384H mutation. This protein features an N-terminal 6xHis-tag, which makes purification straightforward. It achieves a purity greater than 84% as verified by SDS-PAGE. Functional ELISA confirms its biological activity, showing strong binding capabilities with specific antibodies—this suggests it's well-suited for research applications.
SARS-CoV-2's nucleoprotein (N) appears to play a critical role in the viral replication cycle and seems essential for ribonucleoprotein formation. It's involved in packaging the viral RNA genome and modulating host cellular processes. This makes it a key target for research into viral pathogenesis and potential therapeutic interventions. The protein's significance in coronavirus biology studies and vaccine development may underscore its importance in ongoing scientific investigations.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
1. ELISA-Based Antibody Screening and Characterization
The full-length protein with validated binding activity via ELISA supports its use as a capture antigen for detecting anti-nucleoprotein antibodies. The N-terminal 6xHis tag facilitates oriented immobilization. However, the description should note that the Q384H mutation (located in the C-terminal domain) could potentially alter antibody binding epitopes compared to the wild-type protein. Therefore, while suitable for screening, antibodies characterized using this variant may show differential reactivity, and results should be interpreted in the context of this specific mutation.
2. Protein-Protein Interaction Studies
While the protein's biological activity supports interaction studies, the Q384H mutation is a critical factor. The C-terminal domain (where Q384 is located) is involved in protein dimerization and RNA binding. The Q384H mutation could potentially alter interactions with host proteins or viral RNA. Therefore, any interaction data derived from this protein is specific to the Q384H variant and may not represent the wild-type protein's behavior. Studies should be explicitly framed as investigating the impact of the Q384H mutation on interactions.
3. Biochemical and Biophysical Characterization
The protein can be used for biophysical analysis (e.g., circular dichroism, dynamic light scattering) to study its folding and stability. The key application here is the direct comparison with wild-type nucleoprotein to understand the specific effects of the Q384H mutation on structural properties and oligomerization. The purity level (>84%) is acceptable for such techniques, though higher purity is often desirable for some structural methods. The value lies in the comparative analysis enabled by the defined point mutation.
4. Immunoassay Development and Validation
The combination of full-length sequence, His-tag, and, most importantly, ELISA-validated binding activity makes this protein well-suited as a standard or capture antigen in developing and validating quantitative immunoassays (e.g., ELISA) for research purposes. It provides a defined reagent for assay optimization.
5. Antigen Presentation and Immunological Studies
The full-length protein can be used in in vitro systems to study antigen processing and presentation. However, the Q384H mutation could potentially create, destroy, or alter T-cell epitopes, affecting the resulting immune recognition patterns. Therefore, findings on antigen presentation and T-cell responses are specific to this variant. The primary utility is for comparative studies against the wild-type protein to assess the immunologic impact of the Q384H mutation, rather than for defining universal properties of the SARS-CoV-2 nucleoprotein.
Final Recommendation & Action Plan:
This recombinant SARS-CoV-2 nucleoprotein (Q384H) is a valuable tool primarily for immunoassay development and for comparative studies investigating the functional and immunological consequences of the Q384H mutation. The confirmed binding activity via ELISA underscores its reliability for antibody-based applications. For all other uses, especially protein-protein interaction and immunological studies, it is crucial to use this protein specifically to understand the Q384H variant's effects by always including wild-type nucleoprotein controls in experiments. Given the protein's 84% purity, performing additional quality control (e.g., SEC-HPLC) to assess aggregation before sensitive experiments is advisable. Ultimately, researchers should clearly communicate that all data generated is specific to the Q384H variant.
-SDS.jpg)
-AC1.jpg)
-AC2.jpg)
