Ultrahigh-Throughput Single Emulsion Droplet Screening for the Discovery of New B Antigen Cleaving Enzymes
Bibliographic record
Abstract
In search of efficient α-galactosidases that can convert B red blood cells (RBCs) to universal type RBCs, we have developed a simple and robust system for ultrahigh-throughput droplet-based microfluidic screening. Here, a multienzyme coupled assay with a fluorogenic B antigen tetrasaccharide substrate is encapsulated within single emulsion water-in-oil droplets alongside single cells from metagenomic libraries. The resulting fluorescent droplets containing candidate B antigen cleaving enzymes are sorted using a commercially available, walk-up droplet sorting instrument before validation, cloning, and characterization of the hits. Using this approach, we identified and characterized an α-1,3-galactosidase ( Pv GH110) from the human gut microbiome capable of converting B-to-O-type RBCs. The simplicity, efficiency, and accessibility of our microfluidic-based system make it suitable for nonspecialist laboratories and offer a promising tool to discover enzymes that enable the generation of a universal O blood type.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| 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) | 0.000 | 0.000 |
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 teacher head, 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".