Features Partner Sites Information LinkXpress
Sign In
Advertise with Us
RANDOX LABORATORIES

Improved Mass Spectrometric Method Presented for Proteomic Analyses

By Labmedica International staff writers
Posted on 25 Mar 2014
Print article
Image: The NanoACQUITY Ultra Performance Liquid Chromatograph (Photo courtesy of Waters Corporation).
Image: The NanoACQUITY Ultra Performance Liquid Chromatograph (Photo courtesy of Waters Corporation).
Mass spectrometry is a highly sensitive method of measurement of cell components or body fluids that has been used for the analysis of chemical and biological materials.

A proteome represents the entire set of proteins expressed by a cell and through analysis of proteomes, it is possible to obtain a comprehensive picture of the proteins and peptides present in cells or body fluids.

Scientists at the Johannes Gutenberg University (Mainz, Germany) have optimized the mass spectrometric workflow which allows the identification and significantly quantifies more proteins than before. They have perfected a relatively new, data-independent technique that facilitates a very accurate and reproducible quantitative analysis. The team focused in particular on developing novel techniques for quantitative proteomic analysis with the aid of so-called ion mobility mass spectrometry. This technique allows not only measuring the mass of a molecule but can also determine its cross section.

The investigators have also managed to enhance the technique known as label-free quantification. This eliminates the need for samples to be labeled in the laboratory before being analyzed, an otherwise complex procedure. Liquid chromatography-ion mobility separation-mass spectrometry (LC-IMS-MS) was performed using HeLa cell lysates. A software program for the integrated analysis of measurement data called ISOQuant has also been developed. This additional analytical dimension renders the technique optimally suited for the comprehensive investigation of highly complex samples.

Nanoscale separation of tryptic peptides was performed with a nanoACQUITY Ultra Performance Liquid Chromatography (UPLC) system (Waters Corporation, Milford, MA, USA) and mass spectrometric analysis of tryptic peptides was performed using a Synapt G2-S HDMS mass spectrometer, also from Waters Corporation. The average intensity of three most intense peptides strongly correlates with the absolute amount of their source protein. ISOQuant implements the quantification method, and additionally allows user-defined approaches, varying the number of peptides used for protein quantification.

Stefan Tenzer, PhD, an immunologist and the lead author of the study said, “We are now able to directly analyze patient samples and specific immune cells without prior cost-intensive preparation.” His colleagues added that the years of work within the technology platform have paid off in terms of a quantum leap forward with regard to the improvement of the technique of proteomic analysis mass spectrometry. The study was published in the March 2014 edition of the journal Nature Methods.

Related Links:

Johannes Gutenberg University
Waters Corporation



Print article

Channels

Clinical Chemistry

view channel
Image: Dried blood spots are used to screen newborns for a number of rare inherited conditions. A new dried blood spot screening test has been developed for Niemann-Pick type C, a rare neurodegenerative condition that usually is not diagnosed until after damage to brain cells has begun (Photo courtesy of Washington University School of Medicine).

Newborn Screening Test Developed For Deadly Neurological Disorder

A newborn screening test has been introduced that identifies infants with Niemann-Pick disease type C, an often fatal condition in which cholesterol builds up and eventually destroys brain cells.... Read more

Genetic Tests

view channel
Image: The BioMek FXP automation workstation (Photo courtesy of Beckman Coulter).

Innovative Gene Testing Technology Finds Cancer Risks

A new method has been developed for identifying mutations and prioritizing variants in breast and ovarian cancer genes, which will not only reduce the number of possible variants for doctors to investigate,... Read more

Pathology

view channel
Image: A photomicrograph of an abnormal Pap smear showing changes consistent with Human Papilloma Virus (HPV) and Mild Dysplasia. Note the irregular perinuclear cytoplasmic clearing, which is the key characteristic of identifying HPV visually on a Pap smear (Photo courtesy of Dr. Cynthia D Booth, MD).

Combining Two Diagnostic Tests Drastically Reduces Cancer Miss Rates

The Papanicolaou (Pap) test is recommended for women between 21 and 65 years old as a screening test for cervical cancer. Women 30 and older who are negative on co-testing may wait as long as five years... Read more

Lab Tech.

view channel
Image: Candida albicans growing on Sabouraud agar (Photo courtesy of Dr. William Kaplan / CDC).

Chemical Sensor Rapidly Detects Fungal Infections

An elevated concentration of D-arabitol, a simple sugar alcohol, in urine, especially compared to that of L-arabitol or creatinine, is indicative of a fungal infection, which can become deadly.... Read more

Industry News

view channel

R-Biopharm and Merck Partner on Companion Diagnostics

R-Biopharm AG (Darmstadt, Germany) and Merck KGaA (Darmstadt, Germany) have entered into an agreement that constitutes their first collaboration, which will initially focus on developing and launching companion diagnostics. Companion diagnostics play a central role in personalized medicine by helping to find specific... Read more
Copyright © 2000-2016 Globetech Media. All rights reserved.