Biology Reference
In-Depth Information
FIGURE 3
Analytical shifting
from the analyte multiplexing to the
sample throughput in the bio-
marker development pipeline.
validation.
133
cantly
improving the sample throughput for targeted
proteomics is to drastically decrease the sample
complexity via highly speci
One direction for signi
with added reference standards are produced,
each spiked digest (representing one particular
human sample) is coded with a sample-speci
c
mass code via chemical derivatization of func-
tional groups on peptides.
133
It is important to
note that chemicals used are without stable
isotopes, thus much less expensive.
2,133
The
decoupled use of stable isotope labeling and
chemical derivatization makes UMRM an
open-source technology to the community; no
special isotope labeled reagents need to be
synthesized. Its acceptance for validating protein
biomarkers, especially in large number of human
samples, is yet to come.
nity-based
sample preparations coupled with fast
LC-MRM-MS or MALDI-MS/MS analysis.
139
Cost-effective af
caf
nity materials are on the wish-
list of the biomarker community to replace the
currently used, costly anti-peptide antibodies.
The other direction for the sample throughput
improvement is proposed in a proof-of-concept
publication which introduces the technology of
ultra-throughput MRM MS, or UMRM MS.
133
This method converts the capability of contem-
porary QqQ instruments for analyzing thou-
sands of preselected peptides in a single
experiment
126
to analyzing a few signature
peptides of later-stage biomarker candidates in
several tens and hundreds of samples in one
experiment.
133
This method adds common stable
isotope labeled reference standards, proteins, or
peptides to each sample for normalizing the
authentic quantity of signature peptides in the
original samples.
2,133
When proteome digests
CONCLUSION
Rapid technology advances in all aspects
of the MS-based quantitative proteomics
platform
d
experiment design, sample prepara-
tion, separation, MS analysis and data process-
ing, database search, and biological annotation
of experimental results
d
allow biomedical and
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