C‐terminal labelling of recombinant proteins using an engineered split‐intein
Bibliographic record
Abstract
Site‐specific labelling of proteins has great potential to aid in the characterization of protein function, three‐dimensional structure, folding behaviour, and interaction with other proteins or small‐molecules. However, new and more efficient labelling methods are needed to satisfy the increasing demand for creating site‐specifically modified proteins. Here, a novel technique is presented to specifically label recombinant proteins at the C‐terminus using a peptide‐protein trans ‐splicing approach. A label is incorporated into a small synthetic peptide, and then transferred onto the C‐terminus of the protein of interest via a stable peptide bond formed by protein trans ‐splicing. We applied this method to introduce fluorescent labels and biotin at the C‐terminus of recombinant proteins, with biotin being potentially useful in creating protein microchips. The gentle chemistry of the trans ‐splicing method also allowed us to label receptor proteins on the surface of eukaryotic cells, indicating a broad applicability of our labelling technique. Research support was provided by CIHR and NSERC.
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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 | 0.001 |
| 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".