Cloning, expression and purification of the cytokine FLT3-ligand for protein interaction studies
Bibliographic record
Abstract
The FLT3-FLT3L receptor complex is responsible for the proliferation and differentiation of hematopoietic progenitor and stem cells, and is involved in several hematological malignancies. A truncated, soluble form of FLT3L was amplified from the full-length sequence by PCR and cloned into a genomic integration and expression vector for Kluyveromyces lactis. Expression and secretion of the 161 amino acid FLT3L was analyzed by SDS-PAGE and Western blot and showed smeared bands around 50kDa consistent with a high level of varying glycosylation as expected. A yield of 100ug/mL of culture was obtained. Purification was performed by Ni-affinity chromatography and the protein was confirmed to be in the elution fraction via mass spectrometry analysis. Purified FLT3L was able to activate RAW264.7 cells after 24hr incubation period causing large vacuoles and a dendritic appearance. FLT3L will be utilized for LARC protein interaction studies to search for interactors that might serve as therapeutic agents in human body fluids and cell culture models of myeloid leukemia.
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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 | 0.001 |
| Science and technology studies | 0.001 | 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) | 0.003 | 0.003 |
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".