Effect of difluoroacetic acid and biological matrices on the development of a liquid chromatography - triple quadrupole mass spectrometry method for determination of intact growth factor proteins.

07:00 EST 13th February 2020 | BioPortfolio

Summary of "Effect of difluoroacetic acid and biological matrices on the development of a liquid chromatography - triple quadrupole mass spectrometry method for determination of intact growth factor proteins."

In biological systems, variable protein expression is a crucial marker for numerous diseases, including cancer. The vast majority of LC - triple quadrupole MS-based quantitative protein assays use bottom-up methodologies, where proteins are subjected to proteolytic cleavage prior to analysis. Here, the effect of difluoroacetic acid and biological matrices on the developement of a multiple reaction monitoring based top-down reversed-phase LC - triple quadrupole MS method for analysis of cancer-related intact proteins was evaluated. Seven growth factors (5.5 - 26.5 kDa; isoelectric points: 4.6 - 9.9) were analyzed on a wide-pore C4 column. The optimized method was performed at 30°C, using a 0.2 mL/min flow rate, a 10 %B/min gradient slope, and 0.05% (v/v) difluoroacetic acid as a mobile phase modifier. The increase of MS sensitivity due to the difluoroacetic acid (estimated limits of detection in biological matrices 1-500 ng/mL) significantly varied for proteins with lower and higher charge state distributions. Matrix effects, as well as the specificity of the method were assessed for variable biological samples and pretreatment methods. This work demonstrates method development to improve the ability to target intact proteins directly by more affordable triple quadrupole MS instrumentation, which could be beneficial in many application fields. This article is protected by copyright. All rights reserved.


Journal Details

This article was published in the following journal.

Name: Journal of separation science
ISSN: 1615-9314


DeepDyve research library

PubMed Articles [33485 Associated PubMed Articles listed on BioPortfolio]

Rapid sample preparation for ganglioside analysis by liquid chromatography mass spectrometry.

Gangliosides (GG) are glycosphingolipids, composed of a ceramide moiety (fatty acid and long chain base) linked to an oligosaccharide chain containing one (or more) molecule of sialic acid. After lipi...

A rapid, sensitive and widely applicable method for quantitative analysis of underivatized amino acids in different biological matrices by ultra-performance liquid chromatography-tandem mass spectrometry.

A rapid, sensitive and widely applicable method for the simultaneous quantitative analysis of 20 underivatized amino acids in different biological matrices, including serum, plasma and tissue homogena...

Liquid chromatography separation of α- and γ-linolenic acid positional isomers with a stationary phase based on covalently immobilized cellulose tris(3,5-dichlorophenylcarbamate).

α-Linolenic acid (ALA) and its most important positional isomer γ-linolenic acid (GLA), are essential fatty acids (vitamin F). Therefore, ALA- and GLA-rich edible oils hold great potential in human ...

Detection of Fungal Jasmonates by Liquid Chromatography Paired with Mass Spectrometry.

Liquid chromatography-mass spectrometry (LC-MS) is one of the most important analytical chemistry techniques for the detection and characterization of biologically active compounds of low abundance-fo...

Chemical and toxicological determination of lamivudine in biological substances.

The purpose of the study is to develop a method for detecting, isolating and quantifying lamivudine in biological substances. Lamivudine was isolated by liquid-liquid and solid phase extraction. The c...

Clinical Trials [10893 Associated Clinical Trials listed on BioPortfolio]

Emulsion Lipid Digestion & Satiety Study - Effect of Physical State and Acid Stability

The purpose of this study is to compare the changes in blood lipids and feelings of satiety after consumption of acid stable or acid unstable oil-in-water emulsions in which the droplets a...

Effect of Biological Therapy on Biomarkers in Patients With Untreated Hepatitis C, Metastatic Melanoma, or Crohn Disease

RATIONALE: Studying samples of blood from patients with cancer, hepatitis C, or Crohn disease in the laboratory may help doctors learn more about changes that occur in DNA and identify bio...

Liquid Biopsies and IMAging for Improved Cancer Care

The recently developed liquid biopsy technology (to obtain and characterize tumour cells and tumour components like Deoxyribonucleic acid (DNA) or Ribonucleic Acid (RNA) from a simple bloo...

Lysophosphatidic Acid Assay in Patients With Ovarian Cancer or Who Are at Risk for Ovarian Cancer

RATIONALE: Screening tests, such as the lysophosphatidic acid assay, may help doctors find cancer cells early and plan better treatment for ovarian cancer. PURPOSE: This clinical trial is...

Association Between Luminal Bile Salt Content and Duodenal Mucosal Integrity in Functional Dyspepsia

Functional dyspepsia (FD) is an extremely common disorder of gastrointestinal function. Recently, impaired duodenal mucosal integrity was reported as a potential pathophysiological mechani...

Medical and Biotech [MESH] Definitions

Fractionation of a vaporized sample as a consequence of partition between a mobile gaseous phase and a stationary phase held in a column. Two types are gas-solid chromatography, where the fixed phase is a solid, and gas-liquid, in which the stationary phase is a nonvolatile liquid supported on an inert solid matrix.

A method of separation of two or more substances by repeated distribution between two immiscible liquid phases that move past each other in opposite directions. It is a form of liquid-liquid chromatography. (Stedman, 25th ed)

Chromatographic techniques in which the mobile phase is a liquid.

Tabular numerical representations of sequence motifs displaying their variability as likelihood values for each possible residue at each position in a sequence. Position-specific scoring matrices (PSSMs) are calculated from position frequency matrices.

Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.

Quick Search

DeepDyve research library

Relevant Topics

Biological Therapy
Biological therapy involves the use of living organisms, substances derived from living organisms, or laboratory-produced versions of such substances to treat disease. Some biological therapies for cancer use vaccines or bacteria to stimulate the body&rs...

Within medicine, nutrition (the study of food and the effect of its components on the body) has many different roles. Appropriate nutrition can help prevent certain diseases, or treat others. In critically ill patients, artificial feeding by tubes need t...

  Bladder Cancer Brain Cancer Breast Cancer Cancer Cervical Cancer Colorectal Head & Neck Cancers Hodgkin Lymphoma Leukemia Lung Cancer Melanoma Myeloma Ovarian Cancer Pancreatic Cancer ...

Searches Linking to this Article