ArticleRapid communications in mass spectrometry : RCM2026
UHPLC-MS/MS Quantification of Human Cytosolic Aldehyde Oxidase: Heat-Assisted In-Solution Protein Denaturation and Digestion.
Article in Rapid communications in mass spectrometry : RCM, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
0 citing papers in PubMed.
No citing paper in PubMed yet.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors.
Funding
Abstract
rationaleThe quality of protein denaturation and digestion is critical in determining the performance of a UHPLC-MS/MS method in absolute protein quantification. Previous UHPLC-MS/MS methods on human aldehyde oxidase (AOX1) protein provided little or no methodological information on protein denaturation and digestion. Therefore, we developed and optimized a sample preparation method for the UHPLC-MS/MS quantification of human cytosolic AOX1.
methodsVarious reagents and conditions in protein denaturation (urea, Rapigest, temperature, heating time, and acetonitrile) and digestion (incubation time) were evaluated. A UHPLC-MS/MS method was developed and validated for quantifying the surrogate peptide
resultsSurrogate peptide yield was improved by our optimized conditions for AOX1 protein denaturation (heating at 80°C for 15 min together with reduction by DL-dithiothreitol, acetonitrile not added during trypsin digestion) and protein digestion (4 h). A solvent composition of 40% water-10% acetonitrile-50% ammonium bicarbonate buffer enhanced peptide solubility and assay linearity and sensitivity. An AOX1-negative cytosol was an appropriate matrix blank for cellular samples. The UPHLC-MS/MS assay validation metrics were within acceptable FDA guidelines, highlighted by peptide stability during sample preparation, a shorter run time (11.5 min), greater sensitivity (7.5 fmol), broader linearity range (7.5-10 000 fmol), allowing for the low femtomole quantification of human cytosolic AOX1.
conclusionsOur novel methodological advances, as highlighted by a high temperature, in-solution protein denaturation protocol and optimal digestion time, solvent composition, and matrix blank, enabled the femtomole quantification of AOX1 protein using a validated UHPLC-MS/MS method with better assay performance than previous methods.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.