Appl Biochem Biotechnol (2013) 170:774786

DOI 10.1007/s12010-013-0238-7

Serum Proteomics in Biomedical Research: A Systematic

Review
Ai-hua Zhang & Hui Sun & Guang-li Yan & Ying Han & Xi-jun Wang

Received: 3 November 2012 / Accepted: 11 April 2013 /

Published online: 23 April 2013
# Springer Science+Business Media New York 2013

Abstract Proteins that are important indicators of physiological or pathological states may
contribute to the early diagnosis of disease, which may provide a basis for identifying the
underlying mechanism of disease development. Serum, contains an abundance of proteins,
offers an easy and inexpensive approach for disease detection and possesses a high potential
to revolutionize the diagnostics. These differentially expressed proteins in serum have
become an important role to monitoring the state for disease. Availability of emerging
proteomic techniques gives optimism that serum can eventually be placed as a biomedium
for clinical diagnostics. Advancements have benefited biomarker research to the point where
serum is now recognized as an excellent diagnostic medium for the detection of disease.
Comprehensive proteome of human serum fluid with high accuracy and availability has the
potential to open new doors for disease biomarker discovery and for disease diagnostics,
providing insights useful for future study. Thus, this review presents an overview of the
value of serum as a credible diagnostic tool, and we aim to summarize the proteomic
technologies currently used for global analysis of serum proteins and to elaborate on the
application of serum proteomics to the discovery of disease biomarkers, and discuss some of
the critical challenges and perspectives for this emerging field.
Keywords Proteomics . Serum . Proteins . Biomarkers . Disease diagnosis . System biology
Abbreviations
MALDI-TOF-MS
CXCL7
SELDI-TOF-MS
TB
PSA
PD
HCC

Matrix-assisted laser desorption/ionization time-of-flight

mass spectrometry
CXC chemokine ligand 7
Surface enhanced laser desorption/ionization time-of-flight
mass spectrometry
Tuberculosis
Prostate specific antigen
Parkinsons disease
Hepatocellular carcinoma

A.-h. Zhang : H. Sun (*) : G.-l. Yan : Y. Han : X.-j. Wang

National TCM Key Lab of Serum Pharmacochemistry, Key Lab of Chinmedomics, and Heilongjiang
University of Chinese Medicine, Key Pharmacometabolomics Platform of Chinese Medicines,
Heping Road 24, Harbin 150040, China
e-mail: casciences@163.com

Appl Biochem Biotechnol (2013) 170:774786

CRC
LC

775

Colorectal cancer
Laryngeal carcinoma

Introduction
Diseases are often discovered in an advanced stage because of the lack of specific
biomarkers. An early molecular diagnosis is therefore of vital importance in order to
increase the survival rate. Currently, the lack of an easy-to-use and inexpensive
sampling method and lack of an accurate and portable platform to facilitate early
disease detection are the major limitations and have seriously hampered the development of clinical diagnostics [1]. A good diagnostic method should have the characteristics of high sensitivity and specificity and meet the requirements of high
throughput, portability, and low cost for subsequent clinical application [2]. In most
cases, the earlier the disease is diagnosed, the more likely it is to be successfully
cured or well controlled, may dramatically reduce the severity of its impact on the
patients life, or prevent and/or delay subsequent complications [3]. Today, the improved proteomic-discovered technologies are turning serum diagnostics into a clinical
reality [4, 5].
Biomarkers are biomolecules that is associated with an increased risk of the disease and
serve as indicators of biological and pathological processes or physiological and pharmacological responses to a drug treatment. Due to the clinical significance, the identification of
disease biomarkers in serum holds great promise for personalized medicine, especially for
disease diagnosis and prognosis [6]. Human serum contains a large array of proteins; it is an
attractive diagnostic fluid because it has several key advantages for disease diagnosis and
prognosis, for example, minimum cost and easy sample collection and processing [7].
Comprehensive analysis of human serum proteome may contribute to the understanding of
pathophysiology and provide a foundation for the recognition of potential biomarkers of
disease [8, 9]. Proteomics is widely envisioned as playing a significant role in the translation
of genomics to clinically useful applications, especially in the areas of diagnostics and
prognostics. Serum proteomic analysis shows great potential as a useful diagnostic tool and
can facilitate monitoring of both disease progression and effects of therapeutic treatments.
These advantages have ensured the widespread use of serum proteomics as a diagnostic tool
in clinical practice, which will have enormous impact [10].
Most clinical chemistry tests available today rely on old technologies, and these tests are
neither sensitive nor specific for any particular disease, and traditional markers only increase
significantly after substantial disease injury [11]. Therefore, more sensitive markers of
disease are eagerly needed, particularly, for the early detection of disease. Proteomics offers
potential advantages that classical diagnostic approaches do not, based on the following
discovery of a suite clinically relevant biomarker that are simultaneously affected by the
disease [12]. Recently, serum proteomics has demonstrated a great potential for biomarker
discovery and validation for various diseases [13, 14]. The future of this field will depend on
further validation of disease specific biomarkers and their incorporation into state-of-the-art,
the assay that is quantitative, specific, rapid, reliable, sensitive, robust, and cost effective for
broad implementation in diagnostic programs. In order to pave the way for the
universal acceptance of proteomics as a clinically relevant diagnostic tool, we summarize the proteomic technologies currently used for global identification and quantification of serum proteins and elaborate the putative biomarkers discovered for a
variety of human diseases.

776

Appl Biochem Biotechnol (2013) 170:774786

Properties of Serum as a Diagnostic Fluid

Serum as a primary carrier of small molecules in the body contains an enormous amount of
information, making it ideal for the early detection of a wide range of diseases. Serum as a
clinical tool has a number of advantages over other biofluids [15]. It can be obtained in large
sampling quantities, and repeat sampling is not a problem. The advantage and development
have dramatically advanced serum-based diagnostics. Human serum proteomics have proven to be a novel approach in the search for protein biomarkers for the detection of diseases
[16]. Therefore, the analysis of serum proteomes emerges into a field of high interest with
the future goal to maintain and improve livestock productivity and welfare. Currently, serum
proteome analysis represents an important field both for diagnosis and monitoring of various
diseases [1719].
Discovery of biomarkers has become a major focus of disease research, which holds
promising future for early detection, diagnosis, monitoring disease recurrence, and therapeutic treatment efficacy to improve long-term survival of patients. Development of standardized methods for collection and storage of patient samples together with standards for
transportation and handling of samples are needed. This requires knowledge about how
sample processing affects proteome analyses, and thereby how nonbiological biased classification errors are avoided. Insenser et al. had analyzed the impact of storage temperature on
the 2D-DIGE profile of human plasma [20]. 2D-DIGE and MS were used to identify the
differences in the proteomic profiles of human plasma samples stored at either 30 or
80 C for 18 months. Results showed that plasma collections stored at 30 C, and not
only those stored at 80 C, may be used for 2D-DIGE analyses without losing the essential
information about highly abundant proteins. This finding expands the applicability of 2DDIGE to the study of human disease by permitting the analysis of human plasma samples
stored at temperatures between 30 and 80 C. Candidate biomarkers are believed to exist
in very low concentrations and comprise less than 1 % of serum proteins and may be highly
labile as well. Therefore, it is imperative to isolate and enrich low-molecular-weight proteins
from complex mixtures for biomarker discovery.
A key challenge in clinics is the identification of sensitive and specific biomarkers for early
detection, prognostic evaluation, and surveillance of disease. Proteome analysis using human
serum is a technological advancement that will enable the discovery of novel biomarkers and
biomarker patterns of various human diseases [21]. The proteomes of serum represent a
potential gold mine of biological and diagnostic information, but challenges such as dynamic
range of protein concentration have hampered efforts to unlock this resource. Although
proteome analysis using serum has potential in disease prevention, early diagnosis and treatment of diseases, and evaluation of pharmacotherapies, this technology is still in its infancy.
Identification of reliable markers that could predict the development of disease in high-risk
populations would allow for intervention strategies that may prevent evolution to definite
disease [2224]. A compelling need exists for the development of technologies that facilitate
and accelerate the discovery of novel protein biomarkers with therapeutic and diagnostic
potential. Thus, investigative studies about serum biomarker probably afford the best opportunity for the discovery of disease biomarkers.

Serum Diagnostics
People are aware of the importance of regular health check-ups; however, most diseases are
not diagnosed until morbid symptoms become apparent in the late phase. To overcome this

Appl Biochem Biotechnol (2013) 170:774786

777

challenge, medical researchers are devoted to finding disease biomarkers that reveal a hidden
lethal threat before the disease becomes complicated. Fortunately, serum is a readily
accessible and informative biofluid, making it ideal for the early detection of a wide range
of diseases. Serum fluid being the mirror of the body is a perfect medium to be explored
for health and disease surveillance. The translational applications and opportunities are
enormous. Serum proteomics is the application of proteomic technologies to improve a
patients clinical outcomes [25]. Since early treatment is associated with improved clinical
results, it is therefore essential to identify new biomarkers with substantial predictive power
to reduce the serious manifestation [26, 27]. Indeed, considerable efforts and progress have
been made over the last few years in the search for novel biomarkers.
As proteomic technologies continue to mature, serum proteomics have great potential for
biomarker research and clinical applications [28]. An ongoing challenge in proteomics
continues to be the analysis of the serum proteome due to the vast number and complexity
of proteins estimated to be present in this biofluid [29]. With advanced instrumentation and
developed refined analytical techniques, serum proteomics is widely envisioned as a useful
and powerful approach for biomarker discovery [30]. A highly promising first step for most
analytical approaches of serum is to deplete as many proteins as possible. With new and
highly sensitive proteomic technologies, the lower level of analytes in serum is no longer a
limitation. Recently, a large number of medically valuable analytes in serum are gradually
unveiled, and some of them represent biomarkers for different diseases including cancer,
autoimmune diseases, viral diseases, bacterial diseases, cardiovascular diseases, and HIV
[3133]. Interest in serum proteomics as a tool for disease diagnosis and a myriad of other
applications continues to expand at a rapid rate [34, 35]. The state-of-the-art of serum
proteomics is in evolution, and a growing number of proteins will be investigated and
discovered. Thus, serum-based diagnostics may offer a robust alternative for clinicians to
use in the near future to make clinical decisions and predict posttreatment outcomes.

Serum Proteomics
Serum proteome is thought to represent a rich source of biomarkers for early stage disease
detection. Such an approach should not only aid in improved diagnostics, but has already
contributed to the identification of complex signatures that may represent disease subgroups,
early diagnostics, and facilitated the analysis of disease. Nevertheless, three major challenges have hindered biomarker discovery: (a) candidate biomarkers exist at extremely low
concentrations in serum; (b) high abundance resident proteins such as albumin mask the rare
biomarkers; (c) biomarkers are rapidly degraded by endogenous and exogenous proteinases
[36]. Proteomic technologies have been used to analyze the serum protein composition of
serum qualitatively and quantitatively. Analysis of these key proteins has become an
important role to monitor the state of biological organisms and is a widely used diagnostic
tool for disease. Proteomics provides potential advantages that classical diagnostic approaches do not, based on the following discovery of a suite of clinically relevant biomarkers
that are simultaneously affected by the disease [37]. In-depth analysis of the serum proteome
is fundamental to understanding the functions of serum proteins and to reveal disease
biomarkers involved in different pathophysiological conditions, with the ultimate goal of
improving patient diagnosis and prognosis.
Protein biomarkers provide the key diagnostic information for the detection of disease,
risk of disease progression, and a patients likely response to drug therapy. Profiling of the
proteins in serum from a disease population can potentially yield valuable clinical

778

Appl Biochem Biotechnol (2013) 170:774786

parameters to be used for diagnosis and prognosis of the disease [38]. Proteome of whole
serum is highly susceptible to a variety of physiological and biochemical processes.
Comprehensive proteome of human serum fluid with high accuracy and availability has
the potential to open new doors for disease biomarker discovery and for disease diagnostics,
providing insights useful for future study. Serum proteomic analysis utilizes high-throughput
analytical technique in order to assay thousands of serum proteins simultaneously. Different
proteomic tools such as 2D-PAGE, 2D-DIGE, SELDI-ToF-MS, protein arrays, iTRAQ, and
MudPIT technology have been used for differential analysis of various serum biological
samples, to better understand the molecular basis of pathogenesis and the validation and
characterization of disease-associated proteins [39, 40].
Nevertheless, serum proteomics is a field in rapid progression that has already developed
beyond initial criticism and is making its way toward important applications and discoveries.
Specifically, there have been an increasing number of reports on the potential clinical
application of proteomics for early detection as well as risk assessment and management
of disease. In the future, serum proteomic applications in the disease field could identify
serum-based biomarkers that are predictors of disease presence or progression, and serum
proteomics could help define the optimal targeted agent and effective dose for each patients
disease. These advances will allow improved new insights into disease etiology and intervention. Emerging as a promising biofocus, serum proteomics will drive serum analyses and
offer great benefits for public health in the long-term.

Recent and Potential Application to Human Disease Detection

Serum is an attractive medium for disease diagnosis because serum testing has several key
advantages including minimum cost, easy sample collection and processing, and possesses
tremendous potential in clinical diagnostics. Serum proteomics is guaranteed to be an easyto-use and powerful diagnostic tool for defining the onset, progression, and prognosis of
human diseases. As proteomic technologies continue to mature, serum proteomics can
enhance the sensitivity and specificity of human disease detection and have great potential
for biomarker research and clinical applications. The goal of these efforts is to identify
proteins that are uniquely correlated with a specific human disease in order to accurately
diagnose and treat the malady.
Cancer has been the disease with the highest cause of death for a decade. This is partly
due to the lack of ideal tumor markers for early diagnosis, which causes cancer found at the
advanced stage where no curative treatment is available. Therefore, development of tumor
markers with higher sensitivity and specificity is waiting to emerge. Recent advances in
serum proteomic technology made it possible to identify the low abundant proteins in the
clinical samples, and thus extensive efforts are now attempted to search for the tumor
markers [41]. Majority of women with ovarian cancer that is the fifth leading cause of
cancer death in women have advanced stage disease at the time of diagnosis and a poor 5year survival rate [42]. The only available biomarker is CA125, which has an unacceptably
low sensitivity and specificity for diagnostic use. Highly sensitive and specific tools to
further optimize early diagnosis and treatment are needed. Hence, screening has been
investigated in the hopes of improving survival by diagnosing ovarian cancer at an earlier
stage. Serum proteomics is a specific method for cancer diagnosis and provides facilities for
the readily reproducible and reliable detection of tumors in early stages [43]. To identify new
proteins with potential diagnostic or prognostic value for the therapy of ovarian cancer,
comparative proteomic analysis of serum from ovarian cancer patients and healthy women

Appl Biochem Biotechnol (2013) 170:774786

779

were performed. Three proteins with differential abundance were found and identified by
mass spectrometry: -1-antitrypsin, apolipoprotein A-IV, and retinol-binding protein [44]. In
a training set analysis, the three most effective biomarkers exhibited 94 % sensitivity at 98 %
specificity, CA125 alone produced 68 % sensitivity, and the combination increased sensitivity to 88 % [45].
Pancreatic cancer, as a highly malignant cancer and the fourth cause of cancer-related
death in world, is characterized by dismal prognosis, due to rapid disease progression, highly
invasive tumor phenotype, and resistance to chemotherapy. A contributory factor to the poor
outcome is the lack of appropriate, sensitive, and specific biomarkers for early diagnosis;
and despite significant advances in treatment of the disease during the past decade, the
survival rate is little improved. Recently, accompanying the development of proteomic
technology, more and more potential biomarkers have appeared and are being reported
[46]. Matsubara et al. had identified a significant decrease of the plasma CXC chemokine
ligand 7 (CXCL7) level in patients with pancreatic cancer, and combination of CA19-9 with
CXCL7 improved the discriminatory power of the former for pancreatic cancer [47].
Diagnosis of pancreatic cancer at an early stage is important for successful treatment and
improving the prognosis of patients. Serum samples were applied to strong anionic exchange
chromatography protein chips for protein profiling by surface enhanced laser SELDI-TOFMS to distinguish pancreatic cancer from noncancer [48]. Sixty-one protein peaks between
2,000 and 30,000 m/z ratios were detected to establish multiple decision classification trees
for differentiating the known disease states. A sensitivity of 0.833 and a specificity of 1.000
were obtained in distinguishing pancreatic cancer from healthy controls and benign pancreatic diseases. These findings suggest that serum profiling may provide a new diagnostic
option for pancreatic cancer and facilitate early detection of the disease.
Currently, there are no early clinical biomarkers for breast cancer [49]. The discovery of
breast cancer associated serum biomarkers is important for early diagnosis, disease mechanism elucidation, and determination of treatment strategy for the disease. A serum proteomic platform including high abundance protein depletion, lectin affinity fractionation, IEF
separation, and LCMS analysis has been applied to discover breast cancer-associated
proteins [50]. The following candidates, thrombospondin-1 and 5, alpha-1B-glycoprotein,
serum amyloid P-component, and tenascin-X, were selected as promising examples of the
use of this platform. This biomarker panel allows accurate discrimination between breast
cancer and healthy individuals. In addition, it could distinguish subgroups of breast cancer
based on patterns of several specific biomarkers. Pietrowskas study established a high
potential of MALDI-ToF-based analyses for the detection of dynamic changes in the serum
proteome related to therapy of breast cancer patients, which revealed the potential applicability of serum proteome-patterns analyses in monitoring the toxicity of therapy [51].
Pietrowska et al. have identified features of serum proteome patterns that were significantly
different between blood samples of healthy individuals and early stage breast cancer patients
[52]. Further validation of biomarkers could potentially facilitate the early diagnosis of
breast cancer as an aid to imaging diagnostics, and this has progressively led toward more
personalized medicine in regard to treatment options.
Tuberculosis (TB) remains to be a major infectious disease throughout the world.
However, there is no commercially available diagnostic test for this disease with acceptable
sensitivity and specificity for routine laboratory use [53]. New diagnosis tests are urgently
needed to address the global TB burden and to improve control programs especially in
resource-limited settings. One of the potential strategies in developing a new diagnostic
method and in improving the TB vaccine involves the identification of novel antigenic
candidates [54]. The model of biomarkers constructed on the three biomarkers generated

780

Appl Biochem Biotechnol (2013) 170:774786

excellent separation between the TB and control groups [55]. The sensitivity was 84.0 %,
and the specificity was 86.0 %. Blind test data indicated a sensitivity of 80.0 % and a
specificity of 84.2 %. The data suggested a potential application of serum proteomics as an
effective technology to profile proteome; and with pattern analysis, a diagnostic model
comprising of three potential biomarkers was indicated to differentiate people with TB and
healthy controls rapidly and precisely.
Prostate cancer is the most common cancer among men worldwide and is the second
leading cause of death. However, many men who develop a prostate tumor never exhibit
symptoms in the early stage of the disease or even before it spreads to other parts of the
body, such as bones and lymph nodes [56, 57]. Therefore, there is a large drive towards
serum proteomic biomarker discovery, and an underlying necessity to discover specific
markers that may serve as molecular targets for clinically relevant prostate cancer are
needed. Current studies have revealed promising biomarkers for use in diagnosis, assessment of prognosis, and targeting treatment of prostate cancer [58, 59]. Proteomics-based
approaches have the potential to provide more insight into the underlying molecular
mechanisms of the disease and also hold great promise for biomarker discovery in prostate
cancer [60]. Two proteins, pigment epithelium-derived factor and zinc-alpha2-glycoprotein,
have undergone extensive validation in serum and tissue samples from the original cohort
and also from a larger independent cohort of patients. Combination of biomarkers with
clinical and demographic data has produced progress toward the goal of both optimal
screening and risk assessment. On the basis of serum proteomic analysis, four novel
candidates, follistatin, chemokine ligand 16, pentraxin 3, and spondin 2, were validated in
the serum of patients with and without prostate cancer [61]. The proteins presented may be
useful as diagnostic, prognostic, or predictive serological markers for prostate cancer. These
results illustrate the potential of serum proteomic platform that strives to bridge the gap
between discovery and validation of biomarkers for the detection of prostate cancer and help
to explore disease from a new perspective.
Serum biomarkers for neurodegenerative diseases are essential to facilitate disease
diagnosis. Parkinsons disease (PD) is a common disease which occurs in aged people with
chronic, progressive, and degenerative character of the central nervous system. Until now,
there is no effective treatment method in PD patients before they show obvious symptoms
for prevention and early diagnosis. In order to find out early disease biomarkers, twodimensional liquid chromatography-tandem mass spectrometry coupled with iTRAQ labeling was employed to quantitatively identify the differentially expressed proteins among the
different disease progress types of PD [62, 63]. It was found that the expression level of eight
proteins which included serotransferrin and clusterin was increased by proteomic techniques. Those proteins may be associated with oxidative stress, mitochondrial dysfunction,
abnormal protein aggregation, and inflammation. The expression level of apolipoprotein A-I
decreased, particularly in the early stage of PD patients. This protein regulated not only the
lipid metabolism in the central nervous system, but also influenced the deposition process of
proteins which are involved in PD.
Hepatic cirrhosis is a life-threatening disease arising from different chronic liver disorders. Chronic hepatitis C is characterized by a highly variable clinical course, with at least
20 % developing liver cirrhosis within 40 years [64]. Only liver biopsy allows a reliable
evaluation of the course of hepatitis C by grading inflammation and staging fibrosis, and
thus serum biomarkers for hepatic fibrosis with high sensitivity and specificity are needed.
Recent evidence from serum proteomics indicates that MFAP-4 as the novel candidate
biomarker with high diagnostic accuracy for prediction of nondiseased liver versus cirrhosis
[65]. Hepatocellular carcinoma (HCC) is the fifth most common cancer, and advanced

Appl Biochem Biotechnol (2013) 170:774786

781

hepatic fibrosis is a major risk factor for HCC. Recent advancements in quantitative and
large-scale proteomic methods could be used to optimize the clinical application of biomarkers [66]. Early diagnosis of HCC and assessment of the stage of hepatic fibrosis can
also contribute to more effective therapeutic interventions and an improve prognosis.
Furthermore, advancements of proteomic techniques contribute not only to the discovery
of clinically useful biomarkers but also in clarifying the molecular mechanisms of disease
pathogenesis by using serum fluid.
Colorectal cancer (CRC) is the third most common cancer worldwide and has poor
prognosis [67]. To identify the proteins involved in colorectal carcinogenesis, 2-DE and
MALDI-TOF/TOF-based proteomic approach were employed to study the differentially
expressed proteins in tumor and adjacent nontumor tissue samples [68]. Of the seven
significantly and consistently altered proteins identified, hnRNP A1 was one of the most
significantly altered proteins, and its overexpression was confirmed using RT-PCR and
western blot analysis. The data suggested that hnRNP A1 may be a potential biomarker
for early diagnosis, prognosis, and monitoring in the therapy of colorectal cancer. The
incidence of esophageal adenocarcinoma is increasing worldwide, but survival remains
poor. Neoadjuvant chemotherapy can improve survival, but prognostic and predictive biomarkers are required. Serum samples collected before and during the treatment of esophageal cancer and noncancer controls were analyzed by SELDI-TOF-MS [69]. Three peaks,
confirmed as apolipoprotein A-I, serum amyloid A, and transthyretin, were associated by
univariate and multivariate analysis with disease-free survival and overall survival. Plasma
proteins can be detected prior to treatment for esophageal cancer that are associated with
outcome and merit testing as prognostic and predictive markers of response to guide
chemotherapy in esophageal cancer. Serum protein fingerprint of patients with laryngeal
carcinoma (LC) was to screen for protein molecules closely related to LC during the onset
and progression of the disease with SELDI-TOF-MS [70]. The findings suggest that
proteomic diagnostic model could distinguish LC patients from controls with a sensitivity
of 92.1 % and a specificity of 91.9 %. The data suggested that proteomic technology could
be used to screen proteins with altered expression levels in the serum of LC patients.
Serum proteomic approaches that could be used for identifying new biomarkers in
gallbladder cancer may be potential molecular targets for early gallbladder cancer diagnostics and therapeutic applications [71]. Serum samples of endometrial carcinoma were first
depleted of high-abundance proteins, labeled with iTRAQ, and then analyzed via twodimensional liquid chromatography and tandem mass spectrometry [72]. Seven proteins,
orosomucoid 1, haptoglobin, SERPINC 1, alpha-1-antichymotrypsin, apolipoprotein A-IV,
inter-alpha-trypsin inhibitor heavy chain H4, and histidine-rich glycoprotein, were found, for
the first time, to be differentially expressed in atypical endometrial hyperplasia. The
differentially expressed proteins may serve as biomarkers in the diagnosis and follow-up
of endometrial hyperplasia and endometrial carcinoma. Lung cancer is the leading cause of
cancer deaths worldwide. New diagnostics are needed to detect early stage lung cancer
because it may be cured with surgery. In a study done by Ostroff et al., proteomic technology
had identified and developed a 12-protein panel that discriminates lung cancer from controls
with 91 % sensitivity and 84 % specificity in cross-validated training and 89 % sensitivity
and 83 % specificity in a separate verification set, with similar performance for early and late
stage lung cancer [73]. It provides a solid foundation to develop tests sorely needed to
identify early stage lung cancer. The identification of serum biomarkers has lead to improvements in the detection and diagnosis of disease, and combinations of these biomarkers
have increased further their sensitivity and specificity [74]. Consequently, these studies
clearly demonstrated that serum contains proteomic signatures that may serve as biomarkers

782

Appl Biochem Biotechnol (2013) 170:774786

for human diseases, and further studies are needed to fully assess the potential clinical value
of this biomarker candidate.

Concluding Remarks and Future Perspectives

Modern medicine has experienced a tremendous explosion in knowledge about disease pathophysiology, gained largely from understanding the molecular biology of human disease.
Recent advances in proteomics now allow for simultaneous identification and quantification
of thousands of unique proteins in serum biological fluid. In particular, proteomic studies
benefits of serum have been used successfully to discover novel markers of a variety conditions.
With the advantages of an easy and cost-effective diagnostic approach, serum shows high
potential for monitoring general health and disease, possess enormous translational values, and
unparalleled opportunities for clinical applications. This article presents an update on the status
of serum diagnostics and delves into their applications to the discovery of biomarkers for
disease detection and therapeutic applications, with an emphasis on specific high-throughput
biomarkers. With advanced instrumentation and developed analytical techniques, proteomics is
widely envisioned as a useful and powerful approach for serum biomarker discovery.
Human serum proteomics has proven to be a novel approach in the search for protein
biomarkers for detection of human diseases. Comprehensive analysis and identification of the
human serum may contribute to the understanding of the pathophysiology and provide a
foundation for the recognition of potential biomarkers of human disease. Moreover, the avenue
of serum diagnostics incorporating proteomic findings will enable us to connect molecular
analytes to monitor therapies, therapeutic outcomes, and finally disease progression. This
should aid the diagnosis, improve stratification of therapy, and identify novel therapeutic targets
for a variety of diseases, all of which will be essential for the development of personalized and
predictive medicine of the future. Future developments in the field of serum diagnostics can
revolutionize our approach to screening, risk assessment, and therapeutic management for a
range of health conditions. We anticipate that in the near future, this approach will hopefully
allow more individualized treatment to be provided before an advanced stage.
Acknowledgments This work was supported by grants from the Key Program of the Natural Science
Foundation of the State (Grant no. 90709019), the National Key Program on the Subject of Drug Innovation
(Grant no. 2009ZX09502-005), the National Specific Program on the Subject of Public Welfare (Grant no.
200807014), the National Program for Key Basic Research Projects in China (Grant no. 2005CB523406), and
the Foundation of Heilongjiang University of Chinese Medicine (Grant no. 201209).
Competing Financial Interests The authors declare no competing financial interests.

References
1. Schwenk, J., Harmel, N., Zolles, G., Bildl, W., Kulik, A., Heimrich, B., Chisaka, O., Jonas, P., Schulte,
U., Fakler, B., & Klcker, N. (2009). Functional proteomics identify cornichon proteins as auxiliary
subunits of AMPA receptors. Science, 323(5919), 13139.
2. Wang, X., Zhang, A., Han, Y., Wang, P., Sun, H., Song, G., Dong, T., Yuan, Y., Yuan, X., Zhang, M., Xie, N.,
Zhang, H., Dong, H., & Dong, W. (2012). Urine metabolomics analysis for biomarker discovery and detection
of jaundice syndrome in patients with liver disease. Molecular & Cellular Proteomics, 11(8), 37080.
3. Gutirrez-Snchez, G., Atwood, J., Kolli, V. S., Roussos, S., & Augur, C. (2012). Initial proteome
analysis of caffeine-induced proteins in Aspergillus tamarii using two-dimensional fluorescence
difference gel electrophoresis. Applied Biochemistry and Biotechnology, 166(8), 206477.

Appl Biochem Biotechnol (2013) 170:774786

783

4. Wildes, D., & Wells, J. A. (2010). Sampling the N-terminal proteome of human blood. Proceedings of the
National Academy of Sciences of the United States of America, 107(10), 45616.
5. Zhang, Y., Guo, B., & Bi, R. (2012). Ovarian cancer: biomarker proteomic diagnosis in progress. Applied
Biochemistry and Biotechnology, 168(4), 9106.
6. Zhang, A., Sun, H., Sun, W., Ye, Y., & Wang, X. (2013). Proteomic identification network analysis of
haptoglobin as a key regulator associated with liver fibrosis. Applied Biochemistry and Biotechnology,
169(3), 83246.
7. Marondedze, C., & Thomas, L. A. (2012). Apple hypanthium firmness: new insights from comparative
proteomics. Applied Biochemistry and Biotechnology, 168(2), 30626.
8. Aivado, M., Spentzos, D., Germing, U., Alterovitz, G., Meng, X. Y., Grall, F., Giagounidis, A. A.,
Klement, G., Steidl, U., Otu, H. H., Czibere, A., Prall, W. C., Iking-Konert, C., Shayne, M., Ramoni, M.
F., Gattermann, N., Haas, R., Mitsiades, C. S., Fung, E. T., & Libermann, T. A. (2007). Serum proteome
profiling detects myelodysplastic syndromes and identifies CXC chemokine ligands 4 and 7 as markers
for advanced disease. Proceedings of the National Academy of Sciences of the United States of America,
104(4), 130712.
9. Yu, C., Xu, C., Xu, L., Yu, J., Miao, M., & Li, Y. (2012). Serum proteomic analysis revealed diagnostic
value of hemoglobin for nonalcoholic fatty liver disease. Journal of Hepatology, 56(1), 2417.
10. Liu, W., Liu, B., Cai, Q., Li, J., Chen, X., & Zhu, Z. (2012). Proteomic identification of serum biomarkers
for gastric cancer using multidimensional liquid chromatography and 2D differential gel electrophoresis.
Clinical Chemical Acta, 413(1314), 1098106.
11. Wang, X., Zhang, A., & Sun, H. (2012). Future perspectives of Chinese medical formulae: chinmedomics
as an effector. OMICS, 16(78), 41421.
12. Chakraborty, C., Pal, S., Doss, C. G., Wen, Z. H., & Lin, C. S. (2012). In silico analysis of COMT, an
important signaling cascade of dopaminergic neurotransmission pathway, for drug development of
Parkinsons disease. Applied Biochemistry and Biotechnology, 167(4), 84560.
13. Zhang, A., Sun, H., Wang, P., & Wang, X. (2013). Salivary proteomics in biomedical research. Clinica
Chimica Acta, 415, 2615.
14. Besson, D., Pavageau, A. H., Valo, I., Bourreau, A., Blanger, A., Eymerit-Morin, C., Moulire, A.,
Chassevent, A., Boisdron-Celle, M., Morel, A., Solassol, J., Campone, M., Gamelin, E., Barr, B.,
Coqueret, O., & Guette, C. (2011). A quantitative proteomic approach of the different stages of colorectal
cancer establishes OLFM4 as a new nonmetastatic tumor marker. Molecular & Cellular Proteomics,
10(12), M111.009712.
15. Karthik, D., Ilavenil, S., Kaleeswaran, B., Sunil, S., & Ravikumar, S. (2012). Proteomic analysis of
plasma proteins in diabetic rats by 2D electrophoresis and MALDI-TOF-MS. Applied Biochemistry and
Biotechnology, 166(6), 150719.
16. Carlsson, A., Wuttge, D. M., Ingvarsson, J., Bengtsson, A. A., Sturfelt, G., Borrebaeck, C. A., &
Wingren, C. (2011). Serum protein profiling of systemic lupus erythematosus and systemic sclerosis
using recombinant antibody microarrays. Molecular & Cellular Proteomics, 10(5), M110.005033.
17. Wang, X., Zhang, A., Wang, P., Sun, H., Wu, G., Sun, W., Lv, H., Jiao, G., Xu, H., Yuan, Y., Liu, L., Zou,
D., Wu, Z., Han, Y., Yan, G., Dong, W., Wu, F., Dong, T., Yu, Y., Zhang, S., Wu, X., Tong, X., & Meng,
X. (2013). Metabolomics coupled with proteomics advancing drug discovery towards more agile development of targeted combination therapies. Molecular & Cellular Proteomics. doi:10.1074/
mcp.M112.021683.
18. Sun, H., Zhang, A., Yan, G., Han, Y., Sun, W., Ye, Y., & Wang, X. (2013). Proteomics study on
the hepatoprotective effects of traditional Chinese medicine formulae Yin-Chen-Hao-Tang by a
combination of two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser
desorption/ionization-time of flight mass spectrometry. Journal of Pharmaceutical and Biomedical
Analysis, 75, 1739.
19. Wang, X., Zhang, A., Sun, H., Wu, G., Sun, W., & Yan, G. (2012). Network generation enhances
interpretation of proteomics data sets by a combination of two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Analyst,
137(20), 470311.
20. Zhang, K., Yuan, K., Wu, H., Li, Q., Wang, Y., Chen, S., Zhang, L., Gu, H., & Fu, R. (2012).
Identification of potential markers related to neoadjuvant chemotherapy sensitivity of breast cancer by
SELDI-TOF MS. Applied Biochemistry and Biotechnology, 166(3), 75363.
21. Arbing, M. A., Kaufmann, M., Phan, T., Chan, S., Cascio, D., & Eisenberg, D. (2010). The crystal
structure of the Mycobacterium tuberculosis Rv3019c-Rv3020c ESX complex reveals a domain-swapped
heterotetramer. Protein Science, 19(9), 1692703.
22. Mobley, J. A., & Poliakov, A. (2009). Detection of early unfolding events in a dimeric protein by amide
proton exchange and native electrospray mass spectrometry. Protein Science, 18(8), 16207.

784

Appl Biochem Biotechnol (2013) 170:774786

23. Wu, Z., Doondeea, J. B., Gholami, A. M., Janning, M. C., Lemeer, S., Kramer, K., Eccles, S. A., Gollin, S. M.,
Grenman, R., Walch, A., Feller, S. M., & Kuster, B. (2011). Quantitative chemical proteomics reveals new
potential drug targets in head and neck cancer. Molecular & Cellular Proteomics, 10(12), M111.011635.
24. Wiltzius, J. J., Sievers, S. A., Sawaya, M. R., & Eisenberg, D. (2009). Atomic structures of IAPP (amylin)
fusions suggest a mechanism for fibrillation and the role of insulin in the process. Protein Science, 18(7),
152130.
25. Sugiki, T., Yoshiura, C., Kofuku, Y., Ueda, T., Shimada, I., & Takahashi, H. (2009). High-throughput
screening of optimal solution conditions for structural biological studies by fluorescence correlation
spectroscopy. Protein Science, 18(5), 111520.
26. Edrei, Y., Gross, E., Corchia, N., & Abramovitch, R. (2012). Improved efficacy of a novel anti-angiogenic
drug combination (TL-118) against colorectal-cancer liver metastases; MRI monitoring in mice. British
Journal of Cancer, 107(4), 65866.
27. Vafadar-Isfahani, B., Ball, G., Coveney, C., Lemetre, C., Boocock, D., Minthon, L., Hansson, O., Miles,
A. K., Janciauskiene, S. M., Warden, D., Smith, A. D., Wilcock, G., Kalsheker, N., Rees, R., MatharooBall, B., & Morgan, K. (2012). Identification of SPARC-like 1 protein as part of a biomarker panel for
Alzheimers disease in cerebrospinal fluid. Journal of Alzheimers Disease, 28(3), 62536.
28. Afshar, S., Sawaya, M. R., & Morrison, S. L. (2009). Structure of a mutant human purine nucleoside
phosphorylase with the prodrug, 2-fluoro-2-deoxyadenosine and the cytotoxic drug, 2-fluoroadenine.
Protein Science, 18(5), 110714.
29. Hew, C. S., & Gam, L. H. (2011). Proteome analysis of abundant proteins extracted from the leaf of
Gynura procumbens (Lour.) Merr. Applied Biochemistry and Biotechnology, 165(78), 157786.
30. Cubedo, J., Padr, T., Garca-Moll, X., Pint, X., Cinca, J., & Badimon, L. (2011). Proteomic signature of
Apolipoprotein J in the early phase of new-onset myocardial infarction. Journal of Proteome Research,
10(1), 21120.
31. Hu, S., Loo, J. A., & Wong, D. T. (2006). Human body fluid proteome analysis. Proteomics, 6(23), 632653.
32. Navaglia, F., Fogar, P., Basso, D., Greco, E., Padoan, A., Tonidandel, L., Fadi, E., Zambon, C. F.,
Bozzato, D., Moz, S., Seraglia, R., Pedrazzoli, S., & Plebani, M. (2009). Pancreatic cancer biomarkers
discovery by surface-enhanced laser desorption and ionization time-of-flight mass spectrometry. Clinical
Chemistry and Laboratory Medicine, 47(6), 71323.
33. Lim, S. R., Gooi, B. H., Singh, M., & Gam, L. H. (2011). Analysis of differentially expressed proteins in
colorectal cancer using hydroxyapatite column and SDS-PAGE. Applied Biochemistry and Biotechnology,
165(56), 121124.
34. Wang, X., Yang, B., Zhang, A., Sun, H., & Yan, G. (2012). Potential drug targets on insomnia and
intervention effects of Jujuboside. A through metabolic pathway analysis as revealed by UPLC/ESISYNAPT-HDMS coupled with pattern recognition approach. Journal of Proteomics, 75(4), 141127.
35. Zhang, A., Sun, H., Wu, G., Sun, W., Ye, Y., & Wang, X. (2013). Proteomics analysis of hepatoprotective
effects for scoparone using MALDI-TOF/TOF mass spectrometry with bioinformatics. OMICS.
doi:10.1089/omi.2012.0064.
36. Lopez, M. F., Mikulskis, A., Kuzdzal, S., Golenko, E., Petricoin, E. F., 3rd, Liotta, L. A., Patton, W. F.,
Whiteley, G. R., Rosenblatt, K., Gurnani, P., Nandi, A., Neill, S., Cullen, S., OGorman, M., Sarracino,
D., Lynch, C., Johnson, A., Mckenzie, W., & Fishman, D. (2007). A novel, high-throughput workflow for
discovery, and identification of serum carrier protein-bound peptide biomarker candidates in ovarian
cancer samples. Clinical Chemistry, 53(6), 106774.
37. Longo, C., Patanarut, A., George, T., Bishop, B., Zhou, W., Fredolini, C., Ross, M. M., Espina, V.,
Pellacani, G., Petricoin, E. F., 3rd, Liotta, L. A., & Luchini, A. (2009). Core-shell hydrogel particles
harvest, concentrate and preserve labile low abundance biomarkers. PLoS One, 4(3), e4763.
38. Bijian, K., Mlynarek, A. M., Balys, R. L., Jie, S., Xu, Y., Hier, M. P., Black, M. J., Di Falco, M. R.,
LaBoissiere, S., & Alaoui-Jamali, M. A. (2009). Serum proteomic approach for the identification of serum
biomarkers contributed by oral squamous cell carcinoma and host tissue microenvironment. Journal of
Proteome Research, 8(5), 217385.
39. Chen, C. S., Sullivan, S., Anderson, T., Tan, A. C., Alex, P. J., Brant, S. R., Cuffari, C., Bayless, T. M.,
Talor, M. V., Burek, C. L., Wang, H., Li, R., Datta, L. W., Wu, Y., Winslow, R. L., Zhu, H., & Li, X.
(2009). Identification of novel serological biomarkers for inflammatory bowel disease using Escherichia
coli proteome chip. Molecular & Cellular Proteomics, 8(8), 176576.
40. Alterman, M. A., Gogichayeva, N. V., & Kornilayev, B. A. (2004). Matrix-assisted laser desorption/
ionization time-of-flight mass spectrometry-based amino acid analysis. Analytical Biochemistry, 335(2),
18491.
41. Mehan, M. R., Ayers, D., Thirstrup, D., Xiong, W., Ostroff, R. M., Brody, E. N., Walker, J. J., Gold, L.,
Jarvis, T. C., Janjic, N., Baird, G. S., & Wilcox, S. K. (2012). Protein signature of lung cancer tissues.
PLoS One, 7(4), e35157.

Appl Biochem Biotechnol (2013) 170:774786

785

42. Brown, D. L., Andreotti, R. F., Lee, S. I., Dejesus Allison, S. O., Bennett, G. L., Dubinsky, T., Glanc, P.,
Horrow, M. M., Lev-Toaff, A. S., Horowitz, N. S., Podrasky, A. E., Scoutt, L. M., & Zelop, C. M. (2010).
ACR appropriateness criteria ovarian cancer screening. Ultrasound Quarterly, 26(4), 21923.
43. Boyce, E. A., & Kohn, E. C. (2005). Ovarian cancer in the proteomics era: diagnosis, prognosis, and
therapeutic targets. International Journal of Gynecological Cancer, 15(Suppl 3), 26673.
44. Lorkova, L., Pospisilova, J., Lacheta, J., Leahomschi, S., Zivny, J., Cibula, D., Zivny, J., & Petrak, J.
(2012). Decreased concentrations of retinol-binding protein 4 in sera of epithelial ovarian cancer patients:
a potential biomarker identified by proteomics. Oncology Reports, 27(2), 31824.
45. Clarke, C. H., Yip, C., Badgwell, D., Fung, E. T., Coombes, K. R., Zhang, Z., Lu, K. H., & Bast, R. C., Jr.
(2011). Proteomic biomarkers apolipoprotein A1, truncated transthyretin, and connective tissue activating
protein III enhance the sensitivity of CA125 for detecting early stage epithelial ovarian cancer. Gynecologic Oncology, 122(3), 54853.
46. Sun, C., Rosendahl, A. H., Ansari, D., & Andersson, R. (2011). Proteome-based biomarkers in pancreatic
cancer. World Journal of Gastroenterology, 17(44), 484552.
47. Matsubara, J., Honda, K., Ono, M., Tanaka, Y., Kobayashi, M., Jung, G., Yanagisawa, K.,
Sakuma, T., Nakamori, S., Sata, N., Nagai, H., Ioka, T., Okusaka, T., Kosuge, T., Tsuchida, A.,
Shimahara, M., Yasunami, Y., Chiba, T., Hirohashi, S., & Yamada, T. (2011). Reduced plasma
level of CXC chemokine ligand 7 in patients with pancreatic cancer. Cancer Epidemiology,
Biomarkers & Prevention, 20(1), 16071.
48. Guo, J., Wang, W., Liao, P., Lou, W., Ji, Y., Zhang, C., Wu, J., & Zhang, S. (2009). Identification of serum
biomarkers for pancreatic adenocarcinoma by proteomic analysis. Cancer Science, 100(12), 2292301.
49. Bhm, D., Keller, K., Wehrwein, N., Lebrecht, A., Schmidt, M., Klbl, H., & Grus, F. H. (2011). Serum
proteome profiling of primary breast cancer indicates a specific biomarker profile. Oncology Reports,
26(5), 10516.
50. Zeng, Z., Hincapie, M., Pitteri, S. J., Hanash, S., Schalkwijk, J., Hogan, J. M., Wang, H., & Hancock, W.
S. (2011). A proteomics platform combining depletion, multi-lectin affinity chromatography (M-LAC),
and isoelectric focusing to study the breast cancer proteome. Analytical Chemistry, 83(12), 484554.
51. Pietrowska, M., Polanska, J., Marczak, L., Behrendt, K., Nowicka, E., Stobiecki, M., Polanski, A.,
Tarnawski, R., & Widlak, P. (2010). Mass spectrometry-based analysis of therapy-related changes
in serum proteome patterns of patients with early-stage breast cancer. Journal of Translational
Medicine, 8, 66.
52. Pietrowska, M., Marczak, L., Polanska, J., Behrendt, K., Nowicka, E., Walaszczyk, A., Chmura, A., Deja,
R., Stobiecki, M., Polanski, A., Tarnawski, R., & Widlak, P. (2009). Mass spectrometry-based serum
proteome pattern analysis in molecular diagnostics of early stage breast cancer. Journal of Translational
Medicine, 7, 60.
53. Li, Y., Zeng, J., Shi, J., Wang, M., Rao, M., Xue, C., Du, Y., & He, Z. G. (2010). A proteome-scale
identification of novel antigenic proteins in Mycobacterium tuberculosis toward diagnostic and vaccine
development. Journal of Proteome Research, 9(9), 481222.
54. Brust, B., Lecoufle, M., Tuaillon, E., Dedieu, L., Canaan, S., Valverde, V., & Kremer, L. (2011).
Mycobacterium tuberculosis lipolytic enzymes as potential biomarkers for the diagnosis of active
tuberculosis. PLoS One, 6(9), e25078.
55. Liu, J. Y., Jin, L., Zhao, M. Y., Zhang, X., Liu, C. B., Zhang, Y. X., Li, F. J., Zhou, J. M., Wang, H. J., &
Li, J. C. (2011). New serum biomarkers for detection of tuberculosis using surface-enhanced laser
desorption/ionization time-of-flight mass spectrometry. Clinical Chemistry and Laboratory Medicine,
49(10), 172733.
56. Yang, S. Y., Adelstein, J., & Kassis, A. I. (2010). Putative molecular signatures for the imaging of prostate
cancer. Expert Review of Molecular Diagnostics, 10(1), 6574.
57. Yocum, A. K., Khan, A. P., Zhao, R., & Chinnaiyan, A. M. (2010). Development of selected reaction
monitoring-MS methodology to measure peptide biomarkers in prostate cancer. Proteomics, 10(19),
350614.
58. Larkin, S. E., Zeidan, B., Taylor, M. G., Bickers, B., Al-Ruwaili, J., Aukim-Hastie, C., & Townsend, P. A.
(2010). Proteomics in prostate cancer biomarker discovery. Expert Review of Proteomics, 7(1), 93102.
59. Steuber, T., OBrien, M. F., & Lilja, H. (2008). Serum markers for prostate cancer: a rational approach to
the literature. European Urology, 54(1), 3140.
60. Byrne, J. C., Downes, M. R., ODonoghue, N., OKeane, C., ONeill, A., Fan, Y., Fitzpatrick, J. M.,
Dunn, M., & Watson, R. W. (2009). 2D-DIGE as a strategy to identify serum markers for the progression
of prostate cancer. Journal of Proteome Research, 8(2), 94257.
61. Sardana, G., Jung, K., Stephan, C., & Diamandis, E. P. (2008). Proteomic analysis of conditioned media
from the PC3, LNCaP, and 22Rv1 prostate cancer cell lines: discovery and validation of candidate
prostate cancer biomarkers. Journal of Proteome Research, 7(8), 332938.

786

Appl Biochem Biotechnol (2013) 170:774786

62. Zhang, X., Yin, X., Yu, H., Liu, X., Yang, F., Yao, J., Jin, H., & Yang, P. (2012). Quantitative proteomic
analysis of serum proteins in patients with Parkinsons disease using an isobaric tag for relative and
absolute quantification labeling, two-dimensional liquid chromatography, and tandem mass spectrometry.
Analyst, 137(2), 4905.
63. Li, Y. H., Wang, J., Zheng, X. L., Zhang, Y. L., Li, X., Yu, S., He, X., & Chan, P. (2011). Matrix-assisted
laser desorption/ionization time-of-flight mass spectrometry combined with magnetic beads for detecting
serum protein biomarkers in Parkinsons disease. European Neurology, 65(2), 10511.
64. Gressner, O. A., Weiskirchen, R., & Gressner, A. M. (2007). Biomarkers of liver fibrosis: clinical
translation of molecular pathogenesis or based on liver-dependent malfunction tests. Clinica Chimica
Acta, 381(2), 10713.
65. Mlleken, C., Sitek, B., Henkel, C., Poschmann, G., Sipos, B., Wiese, S., Warscheid, B., Broelsch, C.,
Reiser, M., Friedman, S. L., Torne, I., Schlosser, A., Klppel, G., Schmiegel, W., Meyer, H. E.,
Holmskov, U., & Sthler, K. (2009). Detection of novel biomarkers of liver cirrhosis by proteomic
analysis. Hepatology, 49(4), 125766.
66. Uto, H., Kanmura, S., Takami, Y., & Tsubouchi, H. (2010). Clinical proteomics for liver disease: a
promising approach for discovery of novel biomarkers. Proteome Sci., 8, 70.
67. Ward, D. G., Suggett, N., Cheng, Y., Wei, W., Johnson, H., Billingham, L. J., Ismail, T., Wakelam, M. J.,
Johnson, P. J., & Martin, A. (2006). Identification of serum biomarkers for colon cancer by proteomic
analysis. British Journal of Cancer, 94(12), 1898905.
68. Ma, Y. L., Peng, J. Y., Zhang, P., Huang, L., Liu, W. J., Shen, T. Y., Chen, H. Q., Zhou, Y. K., Zhang, M.,
Chu, Z. X., & Qin, H. L. (2009). Heterogeneous nuclear ribonucleoprotein A1 is identified as a potential
biomarker for colorectal cancer based on differential proteomics technology. Journal of Proteome
Research, 8(10), 452535.
69. Kelly, P., Paulin, F., Lamont, D., Baker, L., Clearly, S., Exon, D., & Thompson, A. (2012). Pretreatment
plasma proteomic markers associated with survival in esophageal cancer. British Journal of Cancer,
106(5), 95561.
70. Liu, C., Pan, C., Wang, H., & Yong, L. (2011). Effect of surface-enhanced laser desorption/ionization
time-of-flight mass spectrometry on identifying biomarkers of laryngeal carcinoma. Tumor Biology,
32(6), 113945.
71. Tan, Y., Ma, S. Y., Wang, F. Q., Meng, H. P., Mei, C., Liu, A., & Wu, H. R. (2011). Proteomic-based
analysis for identification of potential serum biomarkers in gallbladder cancer. Oncology Reports, 26(4),
8539.
72. Wang, Y. S., Cao, R., Jin, H., Huang, Y. P., Zhang, X. Y., Cong, Q., He, Y. F., & Xu, C. J. (2011). Altered
protein expression in serum from endometrial hyperplasia and carcinoma patients. Journal of Hematology
& Oncology, 4, 15.
73. Ostroff, R. M., Bigbee, W. L., Franklin, W., Gold, L., Mehan, M., Miller, Y. E., Pass, H. I., Rom, W. N.,
Siegfried, J. M., Stewart, A., Walker, J. J., Weissfeld, J. L., Williams, S., Zichi, D., & Brody, E. N. (2010).
Unlocking biomarker discovery: large scale application of aptamer proteomic technology for early
detection of lung cancer. PLoS One, 5(12), e15003.
74. Han, M. H., Hwang, S. I., Roy, D. B., Lundgren, D. H., Price, J. V., Ousman, S. S., Fernald, G. H., Gerlitz,
B., Robinson, W. H., Baranzini, S. E., Grinnell, B. W., Raine, C. S., Sobel, R. A., Han, D. K., & Steinman,
L. (2008). Proteomic analysis of active multiple sclerosis lesions reveals therapeutic targets. Nature,
451(7182), 107681.