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Validation of Recombinant Antibodies Against Human Transcription Factors

2015· article· en· W2889693399 on OpenAlexaff
Carly Griffin, Shane Miersch, Edyta Marcon, Sunandan Banerjee, Jim Wells, Michael Hornsby, Anthony A. Kossiakoff, Shohei Koide, Marcin Paduch, Sachdev S. Sidhu, Jason Moffat

Bibliographic record

VenueThe FASEB Journal · 2015
Typearticle
Languageen
FieldMedicine
TopicMonoclonal and Polyclonal Antibodies Research
Canadian institutionsUniversity of Toronto
Fundersnot available
KeywordsChromatin immunoprecipitationComputational biologyRecombinant DNATranscription factorImmunoprecipitationBiologyCell biologyCell cultureGene expressionPromoterGeneGenetics

Abstract

fetched live from OpenAlex

The Recombinant Antibody Network (RAN) is a pilot project involving 3 integrated automation centres to create renewable, open‐source, high‐quality binding reagents. The main aims of RAN are to: generate rAbs (recombinant antibodies) directed to the ~1500 human transcription factors (TFs) against ~3000 domain targets using high‐throughput selection methods; validate and produce 蠅2 rAbs per target for high performance in applications such as immunofluorescence (IF), western blot, and chromatin‐immunoprecipitation; distribute validation data to our network and the scientific community through a dedicated web portal; and continually improve the efficiency, speed, cost and success. The RAN pipeline progressively identifies clones that bind target domains with high affinity and specificity. Our rAbs should be able to bind and IP full‐length cellular TFs from complex mixtures, and demonstrate specificity between family members. Additional validation steps include over‐expression of the target, knock‐down of the target by RNAi, and altered growth conditions. A key step in the validation process is high‐content screening against a panel of 6 Human cell lines to monitor cellular localization. The rAbs that show nuclear localization by IF are tested by immunoprecipitation‐mass spectrometry for their ability to pull‐down endogenous target. To date, over 1200 rAbs against >250 targets have been monitored by IF, with approximately 25% showing nuclear localization patterns.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not 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.

metaresearch head score (Codex)0.007
metaresearch head score (Gemma)0.003
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Methods · Consensus signal: none
Teacher disagreement score0.007
Threshold uncertainty score0.038

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0070.003
Meta-epidemiology (narrow)0.0020.001
Meta-epidemiology (broad)0.0010.001
Bibliometrics0.0010.001
Science and technology studies0.0010.001
Scholarly communication0.0010.001
Open science0.0020.001
Research integrity0.0010.001
Insufficient payload (model declined to judge)0.0030.003

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.

Opus teacher head0.097
GPT teacher head0.341
Teacher spread0.244 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreMethods

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".

Quick stats

Citations0
Published2015
Admission routes1
Has abstractyes

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