Method development of immunoglobulin G purification from micro-volumes of human serum for untargeted and targeted proteomics-based antibody repertoire studies

Yu Ting Chang, Ming Chu Chang, Yun Jung Tsai, Christine Ferng, Hsi Chang Shih, Ya Po Kuo, Chung Hsuan Chen, I. Lin Tsai

Research output: Contribution to journalArticle

Abstract

Immunoglobulins (Igs) are major serum proteins which play important roles in immunity. Both untargeted and targeted proteomic workflows can be applied to investigate antigen-binding sites and the glycosylation profiles of Igs. For a more-comprehensive picture of IgG from human serum, we developed an IgG purification process and coupled the standardized method to untargeted and targeted proteomic workflows for IgG investigations. Parameters such as the type of purification beads, volume of the bead slurry, incubation conditions, and binding capacities were evaluated in this study. Only 2 μL of human serum was required for each sample. The performance of coupling the purification process to untargeted proteomics in the IgG analysis was evaluated by comparing normalized abundances of IgG subclass-specific peptides with quantification results from an ELISA. Pearson's correlation values were all >0.82. Targeted proteomic workflow was applied to serum samples from patients with autoimmune pancreatitis and from healthy controls, and the results corresponded to clinical findings that IgG4-related peptides/glycopeptides showed higher abundances in the diseased group. The developed IgG purification process is simple and requires small sample volume, and it can be coupled to targeted and untargeted proteomic workflows for clinical investigations in the future.

Original languageEnglish
JournalJournal of Food and Drug Analysis
DOIs
Publication statusAccepted/In press - Jan 1 2018

Fingerprint

immunoglobulin G
Proteomics
proteomics
Immunoglobulin G
purification methods
Workflow
antibodies
Antibodies
Serum
immunoglobulins
peptides
glycopeptides
methodology
Immunoglobulins
pancreatitis
binding capacity
glycosylation
sampling
blood proteins
Peptides

Keywords

  • Human serum
  • IgG purification
  • Liquid chromatography-mass spectrometry
  • Proteomics

ASJC Scopus subject areas

  • Food Science
  • Pharmacology

Cite this

Method development of immunoglobulin G purification from micro-volumes of human serum for untargeted and targeted proteomics-based antibody repertoire studies. / Chang, Yu Ting; Chang, Ming Chu; Tsai, Yun Jung; Ferng, Christine; Shih, Hsi Chang; Kuo, Ya Po; Chen, Chung Hsuan; Tsai, I. Lin.

In: Journal of Food and Drug Analysis, 01.01.2018.

Research output: Contribution to journalArticle

@article{8a818276d94b45818ece5fb895732387,
title = "Method development of immunoglobulin G purification from micro-volumes of human serum for untargeted and targeted proteomics-based antibody repertoire studies",
abstract = "Immunoglobulins (Igs) are major serum proteins which play important roles in immunity. Both untargeted and targeted proteomic workflows can be applied to investigate antigen-binding sites and the glycosylation profiles of Igs. For a more-comprehensive picture of IgG from human serum, we developed an IgG purification process and coupled the standardized method to untargeted and targeted proteomic workflows for IgG investigations. Parameters such as the type of purification beads, volume of the bead slurry, incubation conditions, and binding capacities were evaluated in this study. Only 2 μL of human serum was required for each sample. The performance of coupling the purification process to untargeted proteomics in the IgG analysis was evaluated by comparing normalized abundances of IgG subclass-specific peptides with quantification results from an ELISA. Pearson's correlation values were all >0.82. Targeted proteomic workflow was applied to serum samples from patients with autoimmune pancreatitis and from healthy controls, and the results corresponded to clinical findings that IgG4-related peptides/glycopeptides showed higher abundances in the diseased group. The developed IgG purification process is simple and requires small sample volume, and it can be coupled to targeted and untargeted proteomic workflows for clinical investigations in the future.",
keywords = "Human serum, IgG purification, Liquid chromatography-mass spectrometry, Proteomics, Human serum, IgG purification, Liquid chromatography-mass spectrometry, Proteomics",
author = "Chang, {Yu Ting} and Chang, {Ming Chu} and Tsai, {Yun Jung} and Christine Ferng and Shih, {Hsi Chang} and Kuo, {Ya Po} and Chen, {Chung Hsuan} and Tsai, {I. Lin}",
year = "2018",
month = "1",
day = "1",
doi = "10.1016/j.jfda.2018.10.001",
language = "English",
journal = "Journal of Food and Drug Analysis",
issn = "1021-9498",
publisher = "Elsevier Taiwan LLC",

}

TY - JOUR

T1 - Method development of immunoglobulin G purification from micro-volumes of human serum for untargeted and targeted proteomics-based antibody repertoire studies

AU - Chang, Yu Ting

AU - Chang, Ming Chu

AU - Tsai, Yun Jung

AU - Ferng, Christine

AU - Shih, Hsi Chang

AU - Kuo, Ya Po

AU - Chen, Chung Hsuan

AU - Tsai, I. Lin

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Immunoglobulins (Igs) are major serum proteins which play important roles in immunity. Both untargeted and targeted proteomic workflows can be applied to investigate antigen-binding sites and the glycosylation profiles of Igs. For a more-comprehensive picture of IgG from human serum, we developed an IgG purification process and coupled the standardized method to untargeted and targeted proteomic workflows for IgG investigations. Parameters such as the type of purification beads, volume of the bead slurry, incubation conditions, and binding capacities were evaluated in this study. Only 2 μL of human serum was required for each sample. The performance of coupling the purification process to untargeted proteomics in the IgG analysis was evaluated by comparing normalized abundances of IgG subclass-specific peptides with quantification results from an ELISA. Pearson's correlation values were all >0.82. Targeted proteomic workflow was applied to serum samples from patients with autoimmune pancreatitis and from healthy controls, and the results corresponded to clinical findings that IgG4-related peptides/glycopeptides showed higher abundances in the diseased group. The developed IgG purification process is simple and requires small sample volume, and it can be coupled to targeted and untargeted proteomic workflows for clinical investigations in the future.

AB - Immunoglobulins (Igs) are major serum proteins which play important roles in immunity. Both untargeted and targeted proteomic workflows can be applied to investigate antigen-binding sites and the glycosylation profiles of Igs. For a more-comprehensive picture of IgG from human serum, we developed an IgG purification process and coupled the standardized method to untargeted and targeted proteomic workflows for IgG investigations. Parameters such as the type of purification beads, volume of the bead slurry, incubation conditions, and binding capacities were evaluated in this study. Only 2 μL of human serum was required for each sample. The performance of coupling the purification process to untargeted proteomics in the IgG analysis was evaluated by comparing normalized abundances of IgG subclass-specific peptides with quantification results from an ELISA. Pearson's correlation values were all >0.82. Targeted proteomic workflow was applied to serum samples from patients with autoimmune pancreatitis and from healthy controls, and the results corresponded to clinical findings that IgG4-related peptides/glycopeptides showed higher abundances in the diseased group. The developed IgG purification process is simple and requires small sample volume, and it can be coupled to targeted and untargeted proteomic workflows for clinical investigations in the future.

KW - Human serum

KW - IgG purification

KW - Liquid chromatography-mass spectrometry

KW - Proteomics

KW - Human serum

KW - IgG purification

KW - Liquid chromatography-mass spectrometry

KW - Proteomics

UR - http://www.scopus.com/inward/record.url?scp=85055870654&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055870654&partnerID=8YFLogxK

U2 - 10.1016/j.jfda.2018.10.001

DO - 10.1016/j.jfda.2018.10.001

M3 - Article

AN - SCOPUS:85055870654

JO - Journal of Food and Drug Analysis

JF - Journal of Food and Drug Analysis

SN - 1021-9498

ER -