Optimized production of the Diels-Alderase antibody 1E9 as a chimeric Fab
Bibliographic record
Abstract
Monoclonal antibody 1E9, which catalyzes the [4+2] cycloaddition between tetrachlorothiophene dioxide and N-ethylmaleimide, has been re-engineered for production as a chimeric humanmurine Fab fragment in Escherichia coli. Stabilizing point mutations in the variable regions of the antibody were identified by replacing residues that rarely occur at individual positions in aligned immunoglobulin sequences with their consensus counterparts. By combining favorable substitutions, double (MetH87ThrGlyL63Ser) and triple (MetH87ThrGlyL63SerPheL95Pro) mutants were created, which can be produced in good yield (4 and 17 mg L1 cell culture, respectively). The triple mutant exhibits a modest fourfold drop in the apparent kcat value for the cycloaddition reaction, but the kinetic properties of the double mutant are indistinguishable from those of the parent murine IgG. The availability of recombinant versions of this catalytic antibody will facilitate efforts to determine the origins of its selectivity and catalytic efficiency through mutagenesis.Key words: catalytic antibody, Fab fragment, bacterial production.
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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".