Small-scale Expression and Purification Protocol for His-SUMO Tagged Enterovirus D68 Strain STL 2014 12 3C Protease in E. coli v1
Bibliographic record
Abstract
This protocol describes the expression and purification of Enterovirus D68 (EV-D68) strain EV-D68 STL 2014 12 3C protease in E. coli Rosetta BL21(DE3)-RR cells. The protein is expressed with an N-terminal His-SUMO tag using an auto-induction system at 18°C. Purification involves cell lysis, initial nickel affinity chromatography (IMAC), SUMO tag removal using His-SENP1 protease, reverse IMAC to remove the cleaved tag, and a final size exclusion chromatography step using a Superdex 75 column. The protocol yields pure protein at approximately 20 mg/mL concentration, suitable for subsequent biochemical and structural studies. The entire process is optimized for small-scale expression (<6L) and takes approximately 5 days from transformation to final purified protein.
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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 | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.001 | 0.003 |
| Insufficient payload (model declined to judge) | 0.008 | 0.015 |
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".