Production and Characterization of Pre-Fusion Ebola Virus GP2 Expressed in Drosophila S2 Cells for Analysis of Vaccine-Induced Immune Responses 3190
Bibliographic record
Abstract
Abstract Description Introduction The Ebola virus (EBOV) glycoprotein (GP) contains two subunits (GP1 and GP2). Upon receptor binding of proteolytically cleaved GP1 in the endosome, pre-fusion GP2 undergoes conformational changes, facilitating membrane fusion to release viral RNA. Our lab has developed a recombinant EBOV GP subunit vaccine with highly protective efficacy in non-human primates (NHPs). However, the mechanism by which GP-specific antibodies confer protection requires further investigation. Our research focuses on investigating the role of GP2-specific antibody binding in vaccine-induced protection. Method: The EBOV GP2 MUT gene was designed incorporating pre-fusion conformation stabilizing mutations and containing a C-tag for purification. Expression plasmids were transfected into Drosophila S2 cells and stably transformed cell lines selected by adding hygromycin. Protein expression was induced by adding CuSO4 and purified using affinity chromatography before characterization by SDS-PAGE and Western blot. Antibody response to GP2 MUT from immunized NHP sera was analyzed using multiplex immunoassay. Results The EBOV GP2 gene was successfully cloned into the pUHM vector, and sequencing confirmed the presence of the targeted mutations and C-tag. GP2 was successfully purified using affinity chromatography and characterized by SDS-PAGE and Western Blot. Stable pre-fusion GP2 was able to detect IgG from immunized NHPs. Funding Sources This project was supported by grants from the Undergraduate Research Opportunities Program, the National Institute of Allergy and Infectious Diseases (R01AI132323), and the National Institute of General Medical Sciences IDeA Networks of Biomedical Research Excellence P20GM103466. The content is solely the responsibility of the authors and does not represent the official views of the NIH. Topic Categories Vaccines and Immunotherapy (VAC)
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".