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

研究成果: 雜誌貢獻文章同行評審

4 引文 斯高帕斯(Scopus)

摘要

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.
原文英語
頁(從 - 到)475-482
頁數8
期刊Journal of Food and Drug Analysis
27
發行號2
DOIs
出版狀態已發佈 - 4月 2019

Keywords

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

ASJC Scopus subject areas

  • 食品科學
  • 藥理

指紋

深入研究「Method development of immunoglobulin G purification from micro-volumes of human serum for untargeted and targeted proteomics-based antibody repertoire studies」主題。共同形成了獨特的指紋。

引用此