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AAAS/Science Business Office FeatureMass Spectrometry
“You have to be aware of what the
limitations are of the instrument, what
you can put into it.”
These days everyone seems to be doing it—mass spectrometry, that is. As
protein chemists, molecular biologists, and even crime scene investigators
rush to adopt this once-esoteric technology, the basic science revolution that
made mass spectrometry user-friendly is striving to make it even more powerful.
By Alan Dove
The revolution inmassspectometry (MS) started inplaces likeCarlos Lebrilla’s
laboratory at theUniversity of California at Davis, California. About 15 years
ago, when the young chemist was a new faculty member at the school,
he hung a sign on his most expensive piece of equipment. The sign, still
hanging today, boldly advertised “Mass Spectrometry for the Masses.” It was not
merely the expression of an inside joke—it was more like a mission statement.
“When I started my career, I saw mass spectrometry going the same way that
NMR went,” says Lebrilla, who is now a full professor. Initially used only by physical
chemists to determine molecular structures, nuclear magnetic resonance (NMR),
rebranded as MRI, is now nearly as ubiquitous in clinical settings as the X-ray. If it
follows thesamepath,massspectrometry,anastonishingly fast, sensitivemethod for
identifying individual molecules in complexmixtures, could revolutionize everything
from clinical diagnostic testing to crime scene investigation. Unfortunately, the
technique has long been considered too expensive, too finicky, and too difficult for
deployment outside of specialized spectroscopy laboratories.
Over the past few years, though, mass spectroscopists and equipment
manufacturers have been chiseling away at those problems, and a new crop of
relatively inexpensive, compact, user-friendly devices is finally bringing the method
within reach of ordinary researchers. But a mass spectrometer is still a substantial
purchase, so scientists considering bringing such a system into their labs should
investigate all of their options first.
Getting Your Charge On
Since J.J. Thomson’s invention of the technique at the turn of the 20th century,
scientists have developed numerous variants of mass spectrometry, but all of them
follow the same essential process. The device draws a sample into a component
called the ionization source, which converts molecules in the sample into ions. The
ions then enter a mass analyzer, which applies different electrostatic and magnetic
forces to the ions in a vacuum, separating them according to their mass-to-charge
(m/z) ratios. Finally, themass analyzer reports these ratios to a connected computer,
which graphs and analyzes the data. From the results, a spectroscopist can identify
specific molecular structures. That is, assuming nothing malfunctions.
“The one thing that anybody who has performed mass spectrometry experiments
is well aware of is mass spectrometers tended to break fairly often,” says Gary
Siuzdak, director of the Scripps Center for Mass Spectrometry in La Jolla, California.
However, he adds that with newer devices, failures in the core components of the
mass spectrometer are relatively rare. Instead, faults seem to now occur more often
in the liquid chromatography (LC) system that is usually coupled to the spectrometer
for sample preparation and sometimes fails due to the complexity and nature of
the samples being injected. “You have to be aware of what the limitations are of
the instrument, what you can put into it,” says Siuzdak, who also wrote
Look for these Upcoming ArticlesProteomics 1—March 7
Biomarker Discovery — March 28
Genomics 1—April 4
continued >
MASS SPECTROMETRY
FOR THE MASSES
Inclusion of companies in this article does not indicateendorsement by either AAAS or Science, nor is it meant toimply that their products or services are superior to those ofother companies.
J.J. Thomson
CREDIT:(TOP)COURTESYOFPACIFIC
NORTHWESTNATIONALLABORATO
RY
LIFE SCIENCE TECHNOLOGIESAAAS/Science Business Office Feature
1116www.sciencemag.org/products
Mass Spectrometry
the standard introductory textbook for biological mass spectrometry
(The Expanding Role of Mass Spectrometry in Biotechnology, 2006).
Even with careful technique, though, the machine could crash,
highlighting one of the key features for which newcomers to mass
spectrometry should shop. “It’s not simply the technology or the
instrument that’s going to be important; it’s going to be the support
structure in getting the instrument installed, training the people on
the instrument and the ongoing challenges,” says Dave Hicks, senior
director of the proteomics business group at Applied Biosystems in
Foster City, California. Like other mass spectrometer makers, Applied
Biosystems provides multiple levels of support with all of its devices,
from field technicians who can fix common glitches to staff scientists
who can answer high-level questions about experimental design and
data analysis.
Besides a deep support network, researchers should look for the
specific features they need, based on the types of samples they’ll
be analyzing. Though there are several kinds of ion sources and
mass analyzers, workers in different fields tend to coalesce around
specific arrangements.
In basic proteomics research, matrix assisted laser desorption/
ionization (MALDI) has become the predominant ion source, and the
mass analyzer is often an ion trap. MALDI is good for ionizing highly
complex samples from protein-friendly buffers, and the ion trap can
eject specific ion species from the sample seriatim, until all of the
ions have been analyzed. It’s a particularly good arrangement for
researchers who aren’t sure what they’re looking for, and need to
capture everything.
Scientists working in a more applied setting, such as a clinical
lab, usually need to process samples more quickly than their basic
research colleagues, but may only need to detect a comparatively
small, defined set of analytes. For these users, electrospray ionization
may be a good choice, combined with a triple-quadrupole mass
analyzer. The triple-quadrupole design isn’t quite as versatile as the
ion trap, but it is efficient and inexpensive.
Less Than a Lamborghini
Of course, the term “inexpensive” is relative. For an entry-level
triple-quadrupole mass spectrometer, complete with a liquid
chromatography system capable of handling a particular class of
samples, expect to spend around $100,000. Adding more features,
more speed, or more versatility can quickly double that figure.
A six-digit purchase will certainly give most researchers pause,
especially in grant-starved academic labs, but the prices on the
latest entry-level mass spectrometers are still a historical bargain.
“No matter what instrument you buy, you’re most likely not going to
lose your job because you bought that instrument,” says Siuzdak. He
adds that a decade ago, when even a cheapmass spectrometer could
cost $300,000 and the technology was less reliable, a poor choice
could indeed shorten one’s career.
Nonetheless, scientistswill stillwant to shoparoundbefore settling
on a particular machine; there are many similar products to choose
from, and each has its own strengths and weaknesses. “I would say
that we’re all competing head to head, but then the practical result is
director of marketing for the LC/MS group at Agilent Technologies in
Santa Clara, California.
Agilent is a relatively recent entrant in the mass spectrometry field,
but the company has quickly developed a large base of users. “I think
we’ve been seeing dramatic growth across whole markets,” says
Miller, adding that the customers seem to be amix of thosemigrating
from other brands and those entering mass spectrometry for the first
time. In response, Agilent has introduced several entry-level models
for different types of users, as well as a series of high-end systems for
experienced spectroscopists.
The same demands have driven longtime mass spectrometer
maker Thermo Fisher Scientific in Waltham, Massachusetts, to
expand its product lines. “Historically one of our key focuses has
been on the high-end systems and the high-end users. But we’re
seeing this transfer of utilization of mass spectrometry to the non-
mass-spec experts,” says Lester Taylor, director of product marketing
for mass spectrometry at the company. Accordingly, Thermo Fisher
has introduced a series of new devices designed for these users.
“There was a strong need to ensure that the operation and use of
these systems is simplified, and that required [revising] the interface
and the software that’s used to run the system,” says Taylor. Indeed,
user-friendly software has become a major selling point for most
LIFE SCIENCE TECHNOLOGIESAAAS/Science Business Office Feature
“It’s not simply the technology
or the instrument that’s going to
be important; it’s going to be the
support structure in getting the
instrument installed, training the
people on the instrument and the
ongoing challenges.”
LIFE SCIENCE TECHNOLOGIESAAAS/Science Business Office Feature
Mac, PC, or MS?
Besides an ionization source and a mass analyzer, every modernmass spectrometer also includes a computer, so it’s no surprise thatdevice makers spend a lot of time developing software. In theory,advanced mass spectroscopists could use other programs to collectand interpret the raw data, but in practice few do.
“Generally I think people use the customized package that comeswith the system from the vendor, because obviously it’s the operatingsystem that drives the instrument, generates the data you need, andis best suited in many applications for processing it,” says LesterTaylor, director of product marketing for mass spectrometry at ThermoFisher Scientific of Waltham, Massachusetts. However, Taylor addsthat many users also export the processed data to other applicationsfor further analysis.
Both experienced and entry-level mass spectroscopists may wantto integrate their mass spectrometry data with other quantitativeexperiments in their labs. Those who already use a laboratoryinformation management package, such as Thermo Fisher’s popularWatsonLIMS, will find that most vendors’ mass spectrometers willexport data to the system fairly easily. In proteomics, researchersoften prefer to send the data to Mascot, a database search tool fromMatrix Science. Mascot uses the spectrometer’s data stream andonline primary protein sequence databases to identify proteins in asample quickly. Researchers can also plug the Mascot data into yetanother application, Proteome Software’s Scaffold, to get even morerefined identification and analysis. Several other software companiesalso offer complementary products.
CREDIT:MACIEJKOTLINSKI
Mass Spectrometry
LIFE SCIENCE TECHNOLOGIESAAAS/Science Business Office Feature
www.sciencemag.org/products 1117
Mass Spectrometry
entry-level mass spectrometers (see “Mac, PC, or MS?”).
While the streamlined interfaces certainly simplify training—
Siuzdak recently taught a high school student to do mass
spectrometry experiments during a rotation in his lab—advances in
the fundamental technology of mass spectrometry have made the
initial purchase more complex. “Even though many people can use
them, you need quite a bit of expertise in helping you pick out the one
that would best suit your needs,” says UC Davis’s Lebrilla. He adds
that while salespeople are a good source for technical information,
prospective buyers should also talk to other researchers in their field
researchers are thinking of buying their first mass spectrometer, but
it won’t be their first exposure to mass spectrometry. “Many of these
sites already have some sort of a core lab or a coremass spectrometry
facility,” Hicks explains, so the question is not whether to use the
technology, but whether to bring it into one’s own laboratory. Users in
this category often need a fairly advanced system, and are probably
less intimidated by complex hardware and software.
When advocates like Siuzdak and Lebrilla talk about bringing the
technology to the “masses,” though, they are usually referring to
userswhohave a specific problem inmind, but little or no background
in mass spectrometry. “We are definitely seeing good growth in these
markets and increasing adoption of mass spectrometry,” says Joe
Anacletto, senior director of applied markets and clinical research at
Applied Biosystems. Unlike Hicks’s customers, Anacletto’s often have
mass spectrometry thrust upon them by regulatory changes.
Environmental testing, food science, and police forensic laborato-
ries exemplify this trend. As the public, legislatures, and courts have
learned more about environmental pollutants and trace evidence,
they have raised their standards for detecting and analyzing specific
compounds. “This drives these testing labs to look for newer tech-
nologies that can handle the broader array of things they’re going to
be looking for,” says Anacletto.
Indeed, the advance of mass spectrometry technology in basic
research labs will likely increase the number of important biological
and environmental markers even further. With the current crop of
robust, sensitive mass spectrometers, biomarker researchers have
embarked on ambitious efforts to find new indicators of disease,
often by comparing blood or serum samples from patients with and
without a particular condition.
Separation Anxiety
Though themass spectrometry has gotten easier in these studies, the
sample preparation problem has been harder to address. Serum, for
example, consists mainly of a handful of highly abundant proteins,
which tend toswampsensitivemassspectrometryassaysandconceal
the much scarcer proteins that might change during pathogenesis.
In response, several companies have introduced immunoaffinity
columns specifically designed to remove these abundant proteins.
The columns, first introduced around 2003 and now available from
companies such as Sigma-Aldrich in St. Louis, Missouri, and Agilent,
have also been refined for greater efficiency and selectivity.
Prefiltering serum samples can certainly simplify a biomarker
study, but the columns also have some disadvantages. Because
they add a step to the sample preparation, they inevitably deplete
some molecules by nonspecific binding. If an important biomarker
is extremely scarce, it might disappear entirely after serum filtration.
Also, scientists hoping to develop assays in animal systems before
moving to human trials may find their protocols hard to translate;
relatively few immunodepletion columns are available for animal
systems, as companies have focused on developing themore popular
human serum columns.
The Matrix and Beyond
Serum is not the only place biomarker investigators have been
searching, of course. For those more interested in characterizing
the markers in solid tissues, a major challenge has been to
determine which cells in a complex sample are actually producing
particular proteins. One elegant option, recently introduced by mass
spectrometry giant Bruker Daltonics of Billerica, Massachusetts, is
an integrated tissue imaging, sample preparation, and spectrometry
platform called ImagePrep. After fixing a layer of tissue to the
system’s stage, researchers can overlay a precisely calibrated matrix,
dry the sample, and performMALDI spectrometry in situ. The result is
a complete picture of the tissue structure, overlaid with spectra of the
proteins expressed in each area.
As the new gear helps generate waves of additional data on every-
thing from biomarkers to environmental chemicals, experts expect
demand to increase for even simpler mass spectrometry platforms,
especially for use in the field or at a patient’s bedside. “The next
stage of that development is to make more portable mass spectrom-
eters. There are a lot of other groups looking at miniaturization of
mass spectrometers, so you can actually just carry it around with
you,” says Lebrilla. Spectrometry for the masses, indeed.
Applied Biosystems
www.appliedbiosystems.com
Agilent Technologies
www.agilent.com
Bruker Daltonics
www.bruker.com
Matrix Science
www.matrixscience.com
Proteome Software
www.proteomesoftware.com
Scripps Center for Mass
Spectrometry
masspec.scripps.edu
Sigma-Aldrich
www.sigma-aldrich.com
Thermo Fisher Scientific
www.thermofisher.com
University of California, Davis
www.ucdavis.edu
Featured Participants
Alan Dove is a science writer and editor based in Massachusetts.
DOI: 10.1126/science.opms.p0800022
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