Integrated analyses of proteins and their glycans in a magnetic bead-based multiplex assay format

Danni Li, Hanching Chiu, Jing Chen, Hui Zhang, Daniel Wan-Yui Chan

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Well-annotated clinical samples are valuable resources for biomarker discovery and validation. Multiplex and integrated methods that simultaneously measure multiple analytes and generate integrated information about these analytes from a single measurement are desirable because these methods help conserve precious samples. We developed a magnetic bead-based system for multiplex and integrated glycoprotein quantification by immunoassays and glycan detection by lectin immunosorbent assays (LISAs). METHODS: Magnetic beads coupled with antibodies were used for capturing proteins of interest. Biotinylated antibodies in combination with streptavidin-labeled phycoerythrin were used for protein quantification. In the LISAs, biotinylated detection antibodies were replaced by biotinylated lectins for glycan detection. RESULTS: Using tissue inhibitor of metallopeptidase 1 (TIMP-1), tissue plasminogen activator, membrane metallo-endopeptidase, and dipeptidyl peptidase-IV (DPP-4) as models, we found that the multiplex integrated system was comparable to single immunoassays in protein quantification and LISAs in glycan detection. The merits of this system were demonstrated when applied to well-annotated prostate cancer tissues for validation of biomarkers in aggressive prostate cancer. Because of the system's multiplex ability, we used only 300 ng of tissue protein for the integrated detection of glycans in these proteins. Fucosylated TIMP-1 and DPP-4 offered improved performance over the proteins in distinguishing aggressive and nonaggressive prostate cancer. CONCLUSIONS: The multiplex and integrated system conserves samples and is a useful tool for validation of glycoproteins and their glycoforms as biomarkers.

Original languageEnglish (US)
Pages (from-to)315-324
Number of pages10
JournalClinical Chemistry
Volume59
Issue number1
DOIs
StatePublished - Jan 2013

Fingerprint

Polysaccharides
Assays
Lectins
Immunosorbents
Biomarkers
Tissue
Prostatic Neoplasms
Proteins
Immunoassay
Antibodies
Glycoproteins
Phycoerythrin
Dipeptidyl Peptidase 4
Neprilysin
Streptavidin
Tissue Plasminogen Activator
metallopeptidase-1

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Integrated analyses of proteins and their glycans in a magnetic bead-based multiplex assay format. / Li, Danni; Chiu, Hanching; Chen, Jing; Zhang, Hui; Chan, Daniel Wan-Yui.

In: Clinical Chemistry, Vol. 59, No. 1, 01.2013, p. 315-324.

Research output: Contribution to journalArticle

@article{c6cfb96b503a4d32a119a435478b2982,
title = "Integrated analyses of proteins and their glycans in a magnetic bead-based multiplex assay format",
abstract = "BACKGROUND: Well-annotated clinical samples are valuable resources for biomarker discovery and validation. Multiplex and integrated methods that simultaneously measure multiple analytes and generate integrated information about these analytes from a single measurement are desirable because these methods help conserve precious samples. We developed a magnetic bead-based system for multiplex and integrated glycoprotein quantification by immunoassays and glycan detection by lectin immunosorbent assays (LISAs). METHODS: Magnetic beads coupled with antibodies were used for capturing proteins of interest. Biotinylated antibodies in combination with streptavidin-labeled phycoerythrin were used for protein quantification. In the LISAs, biotinylated detection antibodies were replaced by biotinylated lectins for glycan detection. RESULTS: Using tissue inhibitor of metallopeptidase 1 (TIMP-1), tissue plasminogen activator, membrane metallo-endopeptidase, and dipeptidyl peptidase-IV (DPP-4) as models, we found that the multiplex integrated system was comparable to single immunoassays in protein quantification and LISAs in glycan detection. The merits of this system were demonstrated when applied to well-annotated prostate cancer tissues for validation of biomarkers in aggressive prostate cancer. Because of the system's multiplex ability, we used only 300 ng of tissue protein for the integrated detection of glycans in these proteins. Fucosylated TIMP-1 and DPP-4 offered improved performance over the proteins in distinguishing aggressive and nonaggressive prostate cancer. CONCLUSIONS: The multiplex and integrated system conserves samples and is a useful tool for validation of glycoproteins and their glycoforms as biomarkers.",
author = "Danni Li and Hanching Chiu and Jing Chen and Hui Zhang and Chan, {Daniel Wan-Yui}",
year = "2013",
month = "1",
doi = "10.1373/clinchem.2012.190983",
language = "English (US)",
volume = "59",
pages = "315--324",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "1",

}

TY - JOUR

T1 - Integrated analyses of proteins and their glycans in a magnetic bead-based multiplex assay format

AU - Li, Danni

AU - Chiu, Hanching

AU - Chen, Jing

AU - Zhang, Hui

AU - Chan, Daniel Wan-Yui

PY - 2013/1

Y1 - 2013/1

N2 - BACKGROUND: Well-annotated clinical samples are valuable resources for biomarker discovery and validation. Multiplex and integrated methods that simultaneously measure multiple analytes and generate integrated information about these analytes from a single measurement are desirable because these methods help conserve precious samples. We developed a magnetic bead-based system for multiplex and integrated glycoprotein quantification by immunoassays and glycan detection by lectin immunosorbent assays (LISAs). METHODS: Magnetic beads coupled with antibodies were used for capturing proteins of interest. Biotinylated antibodies in combination with streptavidin-labeled phycoerythrin were used for protein quantification. In the LISAs, biotinylated detection antibodies were replaced by biotinylated lectins for glycan detection. RESULTS: Using tissue inhibitor of metallopeptidase 1 (TIMP-1), tissue plasminogen activator, membrane metallo-endopeptidase, and dipeptidyl peptidase-IV (DPP-4) as models, we found that the multiplex integrated system was comparable to single immunoassays in protein quantification and LISAs in glycan detection. The merits of this system were demonstrated when applied to well-annotated prostate cancer tissues for validation of biomarkers in aggressive prostate cancer. Because of the system's multiplex ability, we used only 300 ng of tissue protein for the integrated detection of glycans in these proteins. Fucosylated TIMP-1 and DPP-4 offered improved performance over the proteins in distinguishing aggressive and nonaggressive prostate cancer. CONCLUSIONS: The multiplex and integrated system conserves samples and is a useful tool for validation of glycoproteins and their glycoforms as biomarkers.

AB - BACKGROUND: Well-annotated clinical samples are valuable resources for biomarker discovery and validation. Multiplex and integrated methods that simultaneously measure multiple analytes and generate integrated information about these analytes from a single measurement are desirable because these methods help conserve precious samples. We developed a magnetic bead-based system for multiplex and integrated glycoprotein quantification by immunoassays and glycan detection by lectin immunosorbent assays (LISAs). METHODS: Magnetic beads coupled with antibodies were used for capturing proteins of interest. Biotinylated antibodies in combination with streptavidin-labeled phycoerythrin were used for protein quantification. In the LISAs, biotinylated detection antibodies were replaced by biotinylated lectins for glycan detection. RESULTS: Using tissue inhibitor of metallopeptidase 1 (TIMP-1), tissue plasminogen activator, membrane metallo-endopeptidase, and dipeptidyl peptidase-IV (DPP-4) as models, we found that the multiplex integrated system was comparable to single immunoassays in protein quantification and LISAs in glycan detection. The merits of this system were demonstrated when applied to well-annotated prostate cancer tissues for validation of biomarkers in aggressive prostate cancer. Because of the system's multiplex ability, we used only 300 ng of tissue protein for the integrated detection of glycans in these proteins. Fucosylated TIMP-1 and DPP-4 offered improved performance over the proteins in distinguishing aggressive and nonaggressive prostate cancer. CONCLUSIONS: The multiplex and integrated system conserves samples and is a useful tool for validation of glycoproteins and their glycoforms as biomarkers.

UR - http://www.scopus.com/inward/record.url?scp=84872043496&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84872043496&partnerID=8YFLogxK

U2 - 10.1373/clinchem.2012.190983

DO - 10.1373/clinchem.2012.190983

M3 - Article

VL - 59

SP - 315

EP - 324

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 1

ER -