Published Study Validates New Protein Enrichment Approach for Low-Abundance Biomarker Detection

Featured In: Supplier News

Tuesday, April 26, 2024

Get daily Bioscience Technology industry top stories and headlines - Sign up now!

newsvine diigo google
slashdot
Share
Loading...

University of Minnesota researchers found that Bio-Rad Laboratories' ProteoMiner protein enrichment kit enhanced identification of changes to low-abundance proteins and detection of post-translationally modified (PTM) proteins in human saliva. These findings offer promise for improving differential proteomic analyses and biomarker studies aimed at identifying disease-specific proteins and their PTM variants in various types of biological samples and fluids. The study was published in the Journal of Proteome Research.

Even when highly sensitive mass spectrometers are used to analyze complex biological samples and bodily fluids, high-abundance proteins obscure the detection of lower-abundance proteins and their post-translational modifications," said Sri Bandhakavi, who led the study at the University of Minnesota in 2010. (Bandhakavi is now a senior scientist at Bio-Rad.) "These lower-abundance proteins and PTMs are often of most interest to researchers, given their association with specific disease or physiological states."

Unlike immunodepletion methods that are sample-specific, the ProteoMiner kit can be used with a variety of samples. ProteoMiner technology consists of millions of unique hexapeptides immobilized on beads; each bead has potentially unique protein-binding properties. Treatment of samples with the ProteoMiner kit causes partial depletion of high-abundance proteins and simultaneous concentration of low-abundance proteins, resulting in dynamic range compression (DRC) of samples.

Study Findings
Bandhakavi and his former colleagues at the University of Minnesota compared saliva samples from healthy women with those with metastatic breast cancer, and processed the samples with and without using the ProteoMiner kit. The researchers' workflow enabled them to identify low-abundance proteins that were affected in breast cancer samples and that retained quantitative accuracy following treatment with the ProteoMiner kit.

"The workflow is readily transferable to diverse samples, signaling the appropriateness of including ProteoMiner technology in differential analyses of clinical samples or cellular lysates," said Bandhakavi. "Given our ability to identify proteins putatively associated with breast cancer, the study further highlights the systemic diagnostic potential of saliva as a noninvasive alternative to serum, long the gold standard, for biomarker discovery."

The researchers also demonstrated that DRC could serve as a general methodology for increasing identification of various protein PTMs of biological interest in complex samples. Knowledge of these PTMs is often critical to understanding a protein's functional state or potential role in disease. Glycosylated proteins were studied due to their large size and linkage to various diseases.

The researchers coupled the ProteoMiner technology with covalent glycopeptide enrichment and tandem mass spectrometry. With DRC, they identified two times more N-linked glycoproteins and their glycosylation sites than without DRC, increasing the known salivary glycoprotein catalog. The authors concluded that ProteoMiner technology should also boost detectability of other biologically interesting PTMs such as phosphorylation.

Source: Bio-Rad Laboratories

Join the Discussion
Rate Article: Average 0 out of 5
register or log in to comment on this article!

0 Comments

Add Comment

Text Only 2000 character limit

Page 1 of 1

Research Exchange

Bringing the Cell Image into Focus

Nov 2 2010

Improvements in transmission electron microscope (TEM) technology increase the power of this imaging tool for the study of cell biology.

Finding a Cure for Spinal Cord Injury with On-Demand LIMS

Oct 25 2010

The Miami Project to Cure Paralysis finds an on-demand laboratory information management system (LIMS) helps to accelerate discovery in its HCS projects.

Saving Cells: Image Processing for Improved Viability, Part II: Iterative Deconvolution

Oct 25 2010

3D light microscopy and deconvolution provide a means to investigate 3D structure, providing near-confocal quality images without the temporal requirements or potentially damaging phototoxicity associated with other 3D imaging technologies. This article is Part II in a series regarding viability, resolution improvement, and measurement in fluorescence imaging. Part I focused on spectral unmixing.

Saving Cells: Image Processing for Improved Viability

Sep 22 2010

This article is Part I of a two-part series regarding viability, resolution improvement, and measurement in fluorescence imaging. Part II will focus on deconvolution.

Evaluation of a New Nano-Type UV-Vis Spectrophotometer

Mar 3

Analysis of one- to four-microliter size samples for nucleic acids has become routine in many life science laboratories. However, until now, available instruments require considerable manipulation of the instrument and sample; some require manually recording the data. The user must typically lower and raise the arm manually, then wipe the sample manually from the target after each analysis. And fiberoptics used in some of these instruments are subject to deterioration.

Production of Recombinant Proteins and Monoclonal Antibodies in Hollow Fiber Bioreactors

Jan 25

While well-understood, robust and convenient, classical batch-style 2-D culture on non-porous supports or 3-D suspension culture in other devices are really not very biologically relevant models. Cell culture conditions can affect the quality of the antibody or protein produced.

Selecting Robots for Use in Drug Discovery and Testing

Dec 6 2010

Drug discovery and testing, with their need for speed, repeatability and verification, are ideally suited to benefit from robot automation. It is therefore not surprising that robots have been at the forefront of automation developments in both these areas.

HP Scalable Network Storage Systems for Life Sciences

Sep 13 2010

Life sciences research today is advancing exponentially, each step bringing us closer to the realization of truly personalized medicine–preventive care and treatments designed specifically for each individual. In the near future, PCPGM healthcare researchers expect to be able to use predictive genetic testing to create custom treatment plans for individuals and deliver dramatic improvements over today’s one-size-fits-all approach. But research capabilities are only part of the equation; current storage and operating capacities must also evolve to accommodate ever-expanding amounts of data before the goal of personalized medicine can be realized.

Using the Tecan Genesis Workstation to Automate a Cytometric Bead Array (CBA) Immunoassay

Mar 11 2010

The poster describe the process involved in automating a Cytometric Bead Array (CBA) immunoassay developed to measure relative concentrations of serum antibodies against Tetanus (TT), Sperm Whale Myoglobin (SWM) and Keyhole Limpet Hemocyanin (KLH) in KLH-immunized volunteers.

Ensuring Quality in Assays Performed with Automated Liquid Handlers

Feb 2 2010

The focus of this presentation is to highlight the need of ensuring quality in important assays performed with automated liquid handlers. Nearly all assays performed within a laboratory are volume-dependent. In turn, all concentrations of biological and chemical components in these assays, as well as the associated dilution protocols, are volume-dependent. Because analyte concentration is volume-dependent, an assay’s results might be falsely interpreted if liquid handler variability and inaccuracies are unknown or if the system(s) go unchecked for a long period.

Inkjet System for Protein Crystallography

Feb 1 2010

X-ray crystallography is used routinely by scientists to obtain the three dimensional structure of a biological molecule of interest.Such information can be used to determine how a pharmaceutical interacts with a protein target and what changes might improve functionality. However, the crystallization of macromolecules still remains a serious hindrance in structural determination despite impressive advances in screening methods and technologies.

Attention Deficit & Hyperactivity in a Drosophila Memory Mutant

Attention Deficit & Hyperactivity in a Drosophila Memory Mutant

Nov 9 2009

Action selection is modulated by external stimuli either directly or via memory retrieval. In a constantly changing environment, animals have evolved attention-like processes to effectively filter the incoming sensory stream. These attention-like processes, in turn, are modulated by memory. The neurobiological nature of how attention, action selection and memory are inter-connected is unknown. We describe here new phenotypes of the memory mutant radish in the fruit fly Drosophila.

Cyclophosphamide as induction therapy for Wegener's granulomatosis and microscopic polyangiitis

Apr 29

The introduction of cyclophosphamide (CyP) as a treatment for Wegener's granulomatosis (WG) and microscopic polyangiitis (MPA) has been among the most significant contributions in vasculitis. Prior to the introduction of CyP, WG was a uniformly fatal disease,...

Sulphilimine cross-links in Goodpasture's disease

Apr 29

The sulphilimine cross-link of the Goodpasture (GP) autoantigen is a novel molecular mechanism (structural constraint) for conferring immune privilege to a site which otherwise is susceptible to structural changes that induce an immunogenic and pathogenic...

Intravenous immunoglobulins in immunodeficiencies: more than mere replacement therapy

Apr 29

Intravenous immunoglobulin (IVIG) is a therapeutic compound prepared from pools of plasma obtained from several thousand healthy blood donors. For more than 20 years, IVIG has been used in the treatment of a wide range of primary and secondary...

Autoimmunity in primary immune deficiency: taking lessons from our patients

Apr 29

The elucidation of the genes leading to selected immune defects has accelerated our understanding of the molecular basis of tolerance in autoimmunity disorders. Mutations in genes of the immune system are known to lead to a catalogue of functional deficits,...

Prokariotic Cell Collection in Denmark

Nov 6 2009

I would like to know about a prokariotic cell collection in Denmark. Is there a cell bank in this country? I need a Lactobacillus strain for a fermentation assay and this information about the bank is very helpful for me.

Request for Entries

Oct 16 2009

Ask the Experts is your chance to get the answers to questions on applications, materials, methods, processes, and technologies. Email you question to bst_web@advantagemedia.com, and the editors of Bioscience Technology will find an appropriate expert to answer it. Watch this space in the future to see the questions your colleagues are posting.

STAY INFORMED: SUBSCRIBE TO

Magazine and E-mail Newsletters

Loading...
E-mail:   

MULTIMEDIA

Video:

Viewing SureFocus Slides

Jun 11 2010

A demonstration of SureFocus Microscope Slides in the review of AFB Smears. SureFocus Slides are a patent-pending breakthrough in tuberculosis detection, as their fluorescent staining circle remains visible during review, Fluorescence Microscopy.

Podcasts:

Allen Institute for Brain Research

Allen Institute for Brain Research

Oct 14 2009

Discussed in this interview are both the mouse brain project and the human cortex project with an emphasis on the importance of these projects to neuroscience research.

Top Stories and Headlines
EVERYDAY!

FREE Email Newsletter

Information: