Mass spectrometric N-glycan analysis of haptoglobin from patient serum samples using a 96-well plate format

Jianhui Zhu, Jing Wu, Haidi Yin, Jorge Marrero, David M. Lubman

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Alterations in glycosylation of serum glycoproteins can provide unique and highly specific fingerprints of malignancy. Our previous mass spectrometric study revealed that the bifucosylation level of serum haptoglobin was distinctly increased in hepatocellular carcinoma (HCC) patients versus liver cirrhosis of all three major etiologies. We have thus developed a method for the analysis of large numbers of serum samples based on a 96-well plate platform for the evaluation of fucosylation changes of serum haptoglobin between HCC versus cirrhosis. Haptoglobin was isolated from the serum of individual patient samples based on an HPLC column immobilized with antihaptoglobin antibody via hydrazide immobilization chemistry. Only 10 μL of serum was required for glycan extraction and processing for MALDI-QIT mass spectrometry analysis using the 96-well plate format. The bifucosylation degrees of haptoglobin in individuals were calculated using a quantitative glycomics method. The MS data confirmed that the bifucosylated tetra-anntenary glycan was upregulated in HCC samples of all etiologies. This study provides a parallel method for processing glycan content for haptoglobin and evaluating detailed changes in glycan structures for a potentially large cohort of clinical serum samples.

Original languageEnglish (US)
Pages (from-to)4932-4939
Number of pages8
JournalJournal of Proteome Research
Volume14
Issue number11
DOIs
StatePublished - Nov 6 2015

Fingerprint

Haptoglobins
Polysaccharides
Serum
Hepatocellular Carcinoma
Glycosylation
Processing
Glycomics
Liver
Mass spectrometry
Glycoproteins
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Dermatoglyphics
Immobilization
Liver Cirrhosis
Mass Spectrometry
Antibodies
Fibrosis
High Pressure Liquid Chromatography

Keywords

  • 96-well plate format
  • glycan
  • haptoglobin
  • hepatocellular carcinoma
  • HPLC
  • MALDI

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Mass spectrometric N-glycan analysis of haptoglobin from patient serum samples using a 96-well plate format. / Zhu, Jianhui; Wu, Jing; Yin, Haidi; Marrero, Jorge; Lubman, David M.

In: Journal of Proteome Research, Vol. 14, No. 11, 06.11.2015, p. 4932-4939.

Research output: Contribution to journalArticle

Zhu, Jianhui ; Wu, Jing ; Yin, Haidi ; Marrero, Jorge ; Lubman, David M. / Mass spectrometric N-glycan analysis of haptoglobin from patient serum samples using a 96-well plate format. In: Journal of Proteome Research. 2015 ; Vol. 14, No. 11. pp. 4932-4939.
@article{98c5588bc04d4412b69c69abb03868e0,
title = "Mass spectrometric N-glycan analysis of haptoglobin from patient serum samples using a 96-well plate format",
abstract = "Alterations in glycosylation of serum glycoproteins can provide unique and highly specific fingerprints of malignancy. Our previous mass spectrometric study revealed that the bifucosylation level of serum haptoglobin was distinctly increased in hepatocellular carcinoma (HCC) patients versus liver cirrhosis of all three major etiologies. We have thus developed a method for the analysis of large numbers of serum samples based on a 96-well plate platform for the evaluation of fucosylation changes of serum haptoglobin between HCC versus cirrhosis. Haptoglobin was isolated from the serum of individual patient samples based on an HPLC column immobilized with antihaptoglobin antibody via hydrazide immobilization chemistry. Only 10 μL of serum was required for glycan extraction and processing for MALDI-QIT mass spectrometry analysis using the 96-well plate format. The bifucosylation degrees of haptoglobin in individuals were calculated using a quantitative glycomics method. The MS data confirmed that the bifucosylated tetra-anntenary glycan was upregulated in HCC samples of all etiologies. This study provides a parallel method for processing glycan content for haptoglobin and evaluating detailed changes in glycan structures for a potentially large cohort of clinical serum samples.",
keywords = "96-well plate format, glycan, haptoglobin, hepatocellular carcinoma, HPLC, MALDI",
author = "Jianhui Zhu and Jing Wu and Haidi Yin and Jorge Marrero and Lubman, {David M.}",
year = "2015",
month = "11",
day = "6",
doi = "10.1021/acs.jproteome.5b00662",
language = "English (US)",
volume = "14",
pages = "4932--4939",
journal = "Journal of Proteome Research",
issn = "1535-3893",
publisher = "American Chemical Society",
number = "11",

}

TY - JOUR

T1 - Mass spectrometric N-glycan analysis of haptoglobin from patient serum samples using a 96-well plate format

AU - Zhu, Jianhui

AU - Wu, Jing

AU - Yin, Haidi

AU - Marrero, Jorge

AU - Lubman, David M.

PY - 2015/11/6

Y1 - 2015/11/6

N2 - Alterations in glycosylation of serum glycoproteins can provide unique and highly specific fingerprints of malignancy. Our previous mass spectrometric study revealed that the bifucosylation level of serum haptoglobin was distinctly increased in hepatocellular carcinoma (HCC) patients versus liver cirrhosis of all three major etiologies. We have thus developed a method for the analysis of large numbers of serum samples based on a 96-well plate platform for the evaluation of fucosylation changes of serum haptoglobin between HCC versus cirrhosis. Haptoglobin was isolated from the serum of individual patient samples based on an HPLC column immobilized with antihaptoglobin antibody via hydrazide immobilization chemistry. Only 10 μL of serum was required for glycan extraction and processing for MALDI-QIT mass spectrometry analysis using the 96-well plate format. The bifucosylation degrees of haptoglobin in individuals were calculated using a quantitative glycomics method. The MS data confirmed that the bifucosylated tetra-anntenary glycan was upregulated in HCC samples of all etiologies. This study provides a parallel method for processing glycan content for haptoglobin and evaluating detailed changes in glycan structures for a potentially large cohort of clinical serum samples.

AB - Alterations in glycosylation of serum glycoproteins can provide unique and highly specific fingerprints of malignancy. Our previous mass spectrometric study revealed that the bifucosylation level of serum haptoglobin was distinctly increased in hepatocellular carcinoma (HCC) patients versus liver cirrhosis of all three major etiologies. We have thus developed a method for the analysis of large numbers of serum samples based on a 96-well plate platform for the evaluation of fucosylation changes of serum haptoglobin between HCC versus cirrhosis. Haptoglobin was isolated from the serum of individual patient samples based on an HPLC column immobilized with antihaptoglobin antibody via hydrazide immobilization chemistry. Only 10 μL of serum was required for glycan extraction and processing for MALDI-QIT mass spectrometry analysis using the 96-well plate format. The bifucosylation degrees of haptoglobin in individuals were calculated using a quantitative glycomics method. The MS data confirmed that the bifucosylated tetra-anntenary glycan was upregulated in HCC samples of all etiologies. This study provides a parallel method for processing glycan content for haptoglobin and evaluating detailed changes in glycan structures for a potentially large cohort of clinical serum samples.

KW - 96-well plate format

KW - glycan

KW - haptoglobin

KW - hepatocellular carcinoma

KW - HPLC

KW - MALDI

UR - http://www.scopus.com/inward/record.url?scp=84946829402&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84946829402&partnerID=8YFLogxK

U2 - 10.1021/acs.jproteome.5b00662

DO - 10.1021/acs.jproteome.5b00662

M3 - Article

VL - 14

SP - 4932

EP - 4939

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 11

ER -