Plasma glycoprotein profiling for colorectal cancer biomarker identification by lectin glycoarray and lectin blot

Yinghua Qiu, Tasneem H. Patwa, Li Xu, Kerby Shedden, David E. Misek, Missy Tuck, Gracie Jin, Mack Ruffin, Danielle K. Turgeon, Sapna Synal, Robert Bresalier, Norman Marcon, Dean E. Brenner, David M. Lubman

Research output: Contribution to journalArticle

141 Citations (Scopus)

Abstract

Colorectal cancer (CRC) remains a major worldwide cause of cancer-related morbidity and mortality largely due to the insidious onset of the disease. The current clinical procedures utilized for disease diagnosis are invasive, unpleasant, and inconvenient; hence, the need for simple blood tests that could be used for the early detection of CRC. In this work, we have developed methods for glycoproteomics analysis to identify plasma markers with utility to assist in the detection of colorectal cancer (CRC). Following immunodepletion of the most abundant plasma proteins, the plasma N-linked glycoproteins were enriched using lectin affinity chromatography and subsequently further separated by nonporous silica reversed-phase (NPS-RP)-HPLC. Individual RP-HPLC fractions were printed on nitrocellulose coated slides which were then probed with lectins to determine glycan patterns in plasma samples from 9 normal, 5 adenoma, and 6 colorectal cancer patients. Statistical tools, including principal component analysis, hierarchical clustering, and Z-statistics analysis, were employed to identify distinctive glycosylation patterns. Patients diagnosed with colorectal cancer or adenomas were shown to have dramatically higher levels of sialylation and fucosylation as compared to normal controls. Plasma glycoproteins with aberrant glycosylation were identified by nano-LC-MS/MS, while a lectin blotting methodology was used to validate proteins with significantly altered glycosylation as a function of cancer progression. The potential markers identified in this study for diagnosis to distinguish colorectal cancer from adenoma and normal include elevated sialylation and fucosylation in complement C3, histidine-rich glycoprotein, and kininogen-1. These potential markers of colorectal cancer were subsequently validated by lectin blotting in an independent set of plasma samples obtained from 10 CRC patients, 10 patients with adenomas, and 10 normal subjects. These results demonstrate the utility of this strategy for the identification of N-linked glycan patterns as potential markers of CRC in human plasma, and may have the utility to distinguish different disease states.

Original languageEnglish (US)
Pages (from-to)1693-1703
Number of pages11
JournalJournal of Proteome Research
Volume7
Issue number4
DOIs
StatePublished - Apr 1 2008

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Tumor Biomarkers
Lectins
Colorectal Neoplasms
Glycoproteins
Glycosylation
Plasmas
Adenoma
Polysaccharides
Kininogens
Plasma (human)
Affinity chromatography
Complement C3
Collodion
Silicon Dioxide
Principal component analysis
Blood Proteins
High Pressure Liquid Chromatography
Blood
Statistics
Hematologic Tests

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

Qiu, Yinghua ; Patwa, Tasneem H. ; Xu, Li ; Shedden, Kerby ; Misek, David E. ; Tuck, Missy ; Jin, Gracie ; Ruffin, Mack ; Turgeon, Danielle K. ; Synal, Sapna ; Bresalier, Robert ; Marcon, Norman ; Brenner, Dean E. ; Lubman, David M. / Plasma glycoprotein profiling for colorectal cancer biomarker identification by lectin glycoarray and lectin blot. In: Journal of Proteome Research. 2008 ; Vol. 7, No. 4. pp. 1693-1703.
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abstract = "Colorectal cancer (CRC) remains a major worldwide cause of cancer-related morbidity and mortality largely due to the insidious onset of the disease. The current clinical procedures utilized for disease diagnosis are invasive, unpleasant, and inconvenient; hence, the need for simple blood tests that could be used for the early detection of CRC. In this work, we have developed methods for glycoproteomics analysis to identify plasma markers with utility to assist in the detection of colorectal cancer (CRC). Following immunodepletion of the most abundant plasma proteins, the plasma N-linked glycoproteins were enriched using lectin affinity chromatography and subsequently further separated by nonporous silica reversed-phase (NPS-RP)-HPLC. Individual RP-HPLC fractions were printed on nitrocellulose coated slides which were then probed with lectins to determine glycan patterns in plasma samples from 9 normal, 5 adenoma, and 6 colorectal cancer patients. Statistical tools, including principal component analysis, hierarchical clustering, and Z-statistics analysis, were employed to identify distinctive glycosylation patterns. Patients diagnosed with colorectal cancer or adenomas were shown to have dramatically higher levels of sialylation and fucosylation as compared to normal controls. Plasma glycoproteins with aberrant glycosylation were identified by nano-LC-MS/MS, while a lectin blotting methodology was used to validate proteins with significantly altered glycosylation as a function of cancer progression. The potential markers identified in this study for diagnosis to distinguish colorectal cancer from adenoma and normal include elevated sialylation and fucosylation in complement C3, histidine-rich glycoprotein, and kininogen-1. These potential markers of colorectal cancer were subsequently validated by lectin blotting in an independent set of plasma samples obtained from 10 CRC patients, 10 patients with adenomas, and 10 normal subjects. These results demonstrate the utility of this strategy for the identification of N-linked glycan patterns as potential markers of CRC in human plasma, and may have the utility to distinguish different disease states.",
author = "Yinghua Qiu and Patwa, {Tasneem H.} and Li Xu and Kerby Shedden and Misek, {David E.} and Missy Tuck and Gracie Jin and Mack Ruffin and Turgeon, {Danielle K.} and Sapna Synal and Robert Bresalier and Norman Marcon and Brenner, {Dean E.} and Lubman, {David M.}",
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Qiu, Y, Patwa, TH, Xu, L, Shedden, K, Misek, DE, Tuck, M, Jin, G, Ruffin, M, Turgeon, DK, Synal, S, Bresalier, R, Marcon, N, Brenner, DE & Lubman, DM 2008, 'Plasma glycoprotein profiling for colorectal cancer biomarker identification by lectin glycoarray and lectin blot', Journal of Proteome Research, vol. 7, no. 4, pp. 1693-1703. https://doi.org/10.1021/pr700706s

Plasma glycoprotein profiling for colorectal cancer biomarker identification by lectin glycoarray and lectin blot. / Qiu, Yinghua; Patwa, Tasneem H.; Xu, Li; Shedden, Kerby; Misek, David E.; Tuck, Missy; Jin, Gracie; Ruffin, Mack; Turgeon, Danielle K.; Synal, Sapna; Bresalier, Robert; Marcon, Norman; Brenner, Dean E.; Lubman, David M.

In: Journal of Proteome Research, Vol. 7, No. 4, 01.04.2008, p. 1693-1703.

Research output: Contribution to journalArticle

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T1 - Plasma glycoprotein profiling for colorectal cancer biomarker identification by lectin glycoarray and lectin blot

AU - Qiu, Yinghua

AU - Patwa, Tasneem H.

AU - Xu, Li

AU - Shedden, Kerby

AU - Misek, David E.

AU - Tuck, Missy

AU - Jin, Gracie

AU - Ruffin, Mack

AU - Turgeon, Danielle K.

AU - Synal, Sapna

AU - Bresalier, Robert

AU - Marcon, Norman

AU - Brenner, Dean E.

AU - Lubman, David M.

PY - 2008/4/1

Y1 - 2008/4/1

N2 - Colorectal cancer (CRC) remains a major worldwide cause of cancer-related morbidity and mortality largely due to the insidious onset of the disease. The current clinical procedures utilized for disease diagnosis are invasive, unpleasant, and inconvenient; hence, the need for simple blood tests that could be used for the early detection of CRC. In this work, we have developed methods for glycoproteomics analysis to identify plasma markers with utility to assist in the detection of colorectal cancer (CRC). Following immunodepletion of the most abundant plasma proteins, the plasma N-linked glycoproteins were enriched using lectin affinity chromatography and subsequently further separated by nonporous silica reversed-phase (NPS-RP)-HPLC. Individual RP-HPLC fractions were printed on nitrocellulose coated slides which were then probed with lectins to determine glycan patterns in plasma samples from 9 normal, 5 adenoma, and 6 colorectal cancer patients. Statistical tools, including principal component analysis, hierarchical clustering, and Z-statistics analysis, were employed to identify distinctive glycosylation patterns. Patients diagnosed with colorectal cancer or adenomas were shown to have dramatically higher levels of sialylation and fucosylation as compared to normal controls. Plasma glycoproteins with aberrant glycosylation were identified by nano-LC-MS/MS, while a lectin blotting methodology was used to validate proteins with significantly altered glycosylation as a function of cancer progression. The potential markers identified in this study for diagnosis to distinguish colorectal cancer from adenoma and normal include elevated sialylation and fucosylation in complement C3, histidine-rich glycoprotein, and kininogen-1. These potential markers of colorectal cancer were subsequently validated by lectin blotting in an independent set of plasma samples obtained from 10 CRC patients, 10 patients with adenomas, and 10 normal subjects. These results demonstrate the utility of this strategy for the identification of N-linked glycan patterns as potential markers of CRC in human plasma, and may have the utility to distinguish different disease states.

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