Dataset Information

Comparision of Nano and Micro-flow LC-MS/MS Based Single Shot Proteomics

ABSTRACT: In this study, we systematically optimized the micro-flow LC-MS/MS system based ssDDA proteomics. Our results show that combination of 2 – 3 h gradient with prosit rescore method identified the highest number of proteins and peptides. There is no benefit to increase column length. A single 3 h run identified about 9,200 proteins from 50 µg of Arabidopsis protein digest. The micro-flow LC-MS/MS system based ssDDA proteomics can be widely used for routine proteomics analysis of large cohort samples. We compared our micro data with the state of art nano LC data.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Yangyang Bian

LAB HEAD: Bernhard Kuster

PROVIDER: PXD024169 | Pride | 2021-07-01

REPOSITORIES: Pride

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Publications

Comprehensive Single-Shot Proteomics with FAIMS on a Hybrid Orbitrap Mass Spectrometer.

Hebert Alexander S AS Prasad Satendra S Belford Michael W MW Bailey Derek J DJ McAlister Graeme C GC Abbatiello Susan E SE Huguet Romain R Wouters Eloy R ER Dunyach Jean-Jacques JJ Brademan Dain R DR Westphall Michael S MS Coon Joshua J JJ

Analytical chemistry 20180718 15

Liquid chromatography (LC) prefractionation is often implemented to increase proteomic coverage; however, while effective, this approach is laborious, requires considerable sample amount, and can be cumbersome. We describe how interfacing a recently described high-field asymmetric waveform ion mobility spectrometry (FAIMS) device between a nanoelectrospray ionization (nanoESI) emitter and an Orbitrap hybrid mass spectrometer (MS) enables the collection of single-shot proteomic data with comparab ...[more]

PMID: 29969236

Publication: 1/2

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Project description:Lymph-node (LN) metastases predict for high recurrence rates in breast cancer patients. Eradication of micro-metastatic tumor cells is the primary goal of adjuvant systemic treatment. Decisions regarding systemic treatment depend largely on primary tumor characteristics rather than on characteristics of their LN metastases. However, it remains unclear to what extent LN metastases, having already metastasized locally, resemble their primary breast tumors and as such will be eradicated by the systemic therapy chosen. In this study we investigated the genetic differences between primary breast cancers and their paired LN metastases using array comparative genomic hybridization analyses on a high resolution 720K Nimblegen platform. Thus far, no metastasis-specific genomic aberrations have been identified. We hypothesized that this is due to low-resolution platforms and lack of stratification on breast cancer subtypes (specifically, triple-negative (TN) versus luminal). Furthermore, we speculated that as TN tumours are known to be more genetically unstable, their LN metastases would show an increase in random copy number aberrations (CNAs). Therefore, we studied 10 primary TN breast tumourM-bM-^@M-^SLN pairs and 10 luminal pairs and found that all LN metastases clustered nearest to their matched tumour except for two. These two were explained by poor hybridization quality and, interestingly, the presence of two histological components in one tumour. We found no significantly altered CNAs between pairs in the whole group, nor when subdivided over subtypes; neither did we find a CNA increase in LN metastases compared to primary tumours within the TN subgroup, suggesting most CNAs are functional and not random. Our findings suggest a strong clonal relationship between primary breast tumours and its LN metastases and support the use of the primary tumor characteristics to guide adjuvant systemic chemotherapy in breast cancer patients, since primary tumors and their subsequent LN metastases seem remarkably similar, at least prior to treatment. The experiment contains 20 paired primary breast cancer samples with their lymph node metastases, analysed on a 720 whole genome CGH array. Additionally, for one sample its two distinct histological subpopulations were analysed on the same platform.

Dataset Information

Comparision of Nano and Micro-flow LC-MS/MS Based Single Shot Proteomics

Publications

Comprehensive Single-Shot Proteomics with FAIMS on a Hybrid Orbitrap Mass Spectrometer.

Similar Datasets

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