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Record W3197241391 · doi:10.21203/rs.3.rs-819042/v1

A Comparison of Blood Plasma Exosome Enrichment Strategies for Proteomic Analysis

2021· preprint· en· W3197241391 on OpenAlexaff
Natalie Turner, Pevindu Abeysinghe, Keith Cheung, Kanchan Vaswani, Jayden Logan, Paweł Sadowski, Murray D. Mitchell

Bibliographic record

VenueResearch Square · 2021
Typepreprint
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicExtracellular vesicles in disease
Canadian institutionsVictoria Park
FundersAustralian Research CouncilMinistry of Business, Innovation and EmploymentDairyNZ
KeywordsChromatographyUltrafiltration (renal)ChemistryBlood proteinsUltracentrifugeMass spectrometryTandem mass spectrometrySize-exclusion chromatographyBiochemistryEnzyme

Abstract

fetched live from OpenAlex

Abstract Proteomic analysis of exosomes (EX) poses a significant challenge. A ‘gold-standard’ method for plasma EX enrichment for downstream proteomic analysis is yet to be established. Our group has performed a comprehensive study of multi-dimensional enrichment methods to determine their efficiency for protein isolation. Methods were evaluated for their capacity to a) successfully isolate and enrich EX from blood plasma, b) minimise the presence of highly abundant plasma proteins, and c) result in the optimum representation of EX proteins by liquid chromatography tandem mass spectrometry (LC-MS/MS). Blood plasma from four animals (Bos taurus) of similar physical attributes and genetics were used. Three methods of EX enrichment were utilised: ultracentrifugation (UC), size-exclusion chromatography (SEC), and ultrafiltration (UF). These enrichment methods were combined to create four groups for methodological evaluation: UC+SEC, UC+SEC+UF, SEC+UC and SEC+UF. UC+SEC yielded the highest number of protein IDs. Plasma protein identification was the least in SEC+UC, but this method yielded the lowest number of protein IDs overall. UC+SEC+UF decreased EX protein ID and did not improve purity compared to UC+SEC. Our data suggest that the method and sequence of EX enrichment strategy impacts protein ID, which may influence the outcome of biomarker discovery studies.

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)0.002
metaresearch head score (Gemma)0.002
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: Empirical · Consensus signal: Empirical
Teacher disagreement score0.002
Threshold uncertainty score0.010

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0020.002
Meta-epidemiology (narrow)0.0010.000
Meta-epidemiology (broad)0.0010.000
Bibliometrics0.0010.001
Science and technology studies0.0000.000
Scholarly communication0.0010.001
Open science0.0010.001
Research integrity0.0010.000
Insufficient payload (model declined to judge)0.0010.001

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.

Opus teacher head0.059
GPT teacher head0.425
Teacher spread0.366 · 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
GenreEmpirical

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

Citations2
Published2021
Admission routes1
Has abstractyes

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