Mass spectrometry can be used for analysis of small molecules, oligonucleotides,
lipids, glycans, proteins, peptides and determining the location of post translational
modifications. It offers accuracy of measurement over a wide molecular weight range,
as small as sub-picomole, sample consumption. Furthermore, any difference between
theoretical and measured mass may provide important information about the primary
structure and post-translational modifications of a therapeutic protein product.
Bio-Synthesis offers a wide range
of analysis
- MALDI-TOF
- MALDI-TOF/TOF
- LC-MALDI
- LC/MS/MS
- Peptide mass fingerprint analysis
- Post-translational modification identification
- Stable isotope related procedures ( iTRAQ, and 18O
and SILAC)
Bio-Synthesis' mass spectrometry services uses industry-leading technologies—instruments,
chemistry, software, and quantification tools -- that are specifically optimized
for exceptional, application-focused performance. This end-to-end system is designed
to deliver the fast and reliable quality control methods for your proteins and peptides.
Please
contact us to discuss your specific project
in more detail. We will provide you with a full project proposal within 2 days that
includes analysis protocols, timelines, and costs.
Identify and
QA your protein/peptide by MS starting at $150
Services and
Fees
Contact us for services and fees not listed below.
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Services
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Cost/Sample
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Electrospray Mass Spectrometry Analysis
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Electrospray Mass Spectra (ESI)
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$150
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Electrospray Mass Spectra (APCI)
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$150
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ES-MS/MS
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$200
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Liquipd Chromatography Mass Spectra (LC-MS)
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$285
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Liquipd Chromatography Mass Spectra (LC-MS/MS)
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$300
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MALDI Mass Spectrometry Analysis
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MALDI-TOF
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$150
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MALDI (Linear=Reflection)
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$244
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MALDI (Digest)
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$215
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MALDI (Tof-Tof of MS/MS)
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$200
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Others
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Desalting
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$70
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Tryspin Digest
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$70
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Phosphorylation Enrichment
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$100
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HPLC PDA (Photo Diode Array UV-vis)
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$150
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HPLC Fluorescence Detection
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$195
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Optical Rotation
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$150
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Proteomic Facility's Major Core
Equipment
- Applied Biosystems Voyager-DE STR MALDI-TOF
- Waters/Micromass Qtof-2 with Waters Cap LC system
- Applied Biosystems 4700 Proteomics Analyzer (MALDI TOF/TOF)
- Applied Biosystems 4800 MALDI TOF/TOF Analyzer
- Applied Biosystems 4000 QTRAP with LC Packings capillary LC
system
Quality Control Data
quality is ensured through the use of appropriate internal and external standards.
The MALDI-TOF mass spectrometers are calibrated with external standards that vary
depending on the analyte molecular weight. This results in the best calibration
for the most accurate mass assignments. Samples that have been trypsin digested
are internally recalibrated from observed trypsin autolysis fragments. The electrospray
mass spectrometers are calibrated using polyalanine which allows for even distribution
of multiple mass peaks. A variety of search engines, for protein identification
from mass fingerprinting and for interpretation of MS/MS spectra, are available
each of which generates its own statistics. The choice of database searching software
to be applied is optional to investigators and discussion of this matter is encouraged
with our technical support team.
Sample Submission
and Ordering
Sample Storage
- Lyophilize or speed vac the protein to a solid sample to ensure
protein stability during shipment. Store sample in Eppendorf Safe Lock tube packed
in such a way that a cushioned from the effects of FedEX. If a gasket-equipped tube
is not available, use Parafilm to assure that the cap does not pop off in shipping.
Keeping the sample cool is at the discretion of the investigator. It is not necessary
to preserve biological activity or structural integrity of the sample unless solubility
will become an issue. Dried samples and samples on PVDF do not require being kept
cool.
- Alternatively, refrigerate to +4°C for cold shipment of the
liquid sample.
Sample Shipment
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1.
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Before sending a sample, please use the mass spectrometry sample submission form
to alert us for the arrival of the sample. Concurrently, a printed copy of the mass
spectrometry sample submission form should accompany the sample. This form is available
on this website and can be filled out online.
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2.
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Ship the sample to:
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Bio-Synthesis Inc
Bioanalytical Laboratory
Attn: Mass Spectrometry
612 E. Main Street
Lewisville, TX 75057
800.227.0627 | 972-420-8505
Mass Spectrometry
Sample Preparation Guidelines
Prior to sending samples, investigators are recommended to contact the facility
to discuss the required analysis. This is necessary to ensure that the most efficient
and cost-effective analytical methods are employed. Samples are normally analyzed
in the order of their receipt, but special arrangements can be made for unstable
samples. A sample submission form and guidelines should accompany each set of samples.
General guidelines for sample preparation:
When preparing samples for mass spectrometric analysis, it is best to limit sample
manipulations. When dealing with solution samples, never bring the sample to complete
dryness. Salts and detergents will interfere with MS analysis of solution samples.
Based on extensive studies, we have found that
SDS-PAGE or 2D IEF-SDS-PAGE is the best method for recovery of low quantities of
protein.
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1.
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The sample should be dissolved in pure water or shipped completely dry if possible.
Try to avoid sending samples in high ionic strength buffers as this may interfere
with the ionization process. Organic buffers, such as tris and guanidine, and inorganic
buffers, such as PBS, are well known for suppressing the ionization process and
should be avoided if possible.
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2.
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If the sample is shipped in solution containing any salt, please provide an estimate
of the salt concentration possible. Also, providing a list of the salts and buffers
that might be present in the solution is also extremely helpful.
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3.
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Any amount of sample can be submitted for analysis. Due to the fact the ionization
process and detector sensitivity are sample dependent, it is rather difficult to
specify a minimum amount of sample necessary for analysis. The general rule of thumb
is, "The more the better".
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Sample amount recommendation:
LC MS/MS
- Deliver more accurate data for larger proteins - MW above 50
kDa.
- Sample amount = 500 attomols of peptide is routinely identified.
- Recommended amount is 1-10 fmol.
- Coomassie blue and digestion compatible silver stained protein
bands are appropriate.
MALDI-MS & - Tof/Tof
- Suitable for intact protein mass analysis up to 250K (mass
resolution expected to be ±250 ppm).
- Analysis of DIGE protein spot digests with a sensitivity of
∼250 attomol routinely.
- Comparative MALDI-MS mapping of control versus experimental
digests.
- MS and MS/MS analysis of beta peptides.
- A variety of biomolecules can be analyzed including oligosaccharides,
and gangliosides.
- Best suited for simple protein mixtures of 1-3 proteins. More
complex mixtures are analyzed by LC-MS/MS using either a single or MudPIT run.
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4.
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A list of information about the sample is usually necessary. This usually includes
such information as sample type (peptide, protein, etc.), expected molecular weight,
purity, etc.
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5.
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Radiolabeled samples may not be submitted for mass analysis.
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Tips:
Protein/peptide solutions in 10 μM volatile buffers can sometimes be analyzed directly
while samples with higher salt concentrations require prior C18 or C4 ZipTip desalting.
Links and Resources
Order and Quote
Mass Spectrometry Online Order Submission Form
Selected Reading
1.
Mann, M., Kelleher, N.L., Precision Proteomics: The case for high resolution and
high mass accuracy, Proc Natl Acad Sci U S A. 2008 Nov 25;105(47):18132-8.
2.
Graumann, J., et. al., SILAC-labeling and proteome quantitation of mouse embryonic
stem cells to a depth of 5111 proteins, Mol Cell Proteomics. 2008 Apr;7(4):672-83.
Epub 2007 Nov 28.
3.
Haas, W., et. al., Optimization and use of peptide mass measurement accuracy in
shotgun proteomics, Mol Cell Proteomics. 2006 Jul;5(7):1326-37. Epub 2006 Apr 23.
4.
Walker, J.M., The Proteomics Protocols Handbook, 2005 Simpson, R.J., Purifying Proteins
for Proteomics: A Laboratory Manual, 2004.
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