ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis.
A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. The workflow involves isolation of the haptoglobin using an HPLC-based affinity column followed by glycan removal, extraction, and desialylation. The fucosylated glycans are then derivatized by Meladrazine, which significantly enhances the detection of the glycans in electrospray ionization. The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC.
Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.|Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
Yiwei
Y
Zhang
Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
Jianhui
J
Zhu
Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
Haidi
H
Yin
Liver Transplantation Program, University of Texas Southwestern Medical Center , Dallas, Texas 75390, United States.
Jorge
J
Marrero
Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
Xin-Xiang
XX
Zhang
Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
David M
DM
Lubman
D014408:Biomarkers, Tumor / Q000097:blood / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005643:Fucose / Q000097:blood; D006031:Glycosylation; D006242:Haptoglobins / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D008103:Liver Cirrhosis / Q000097:blood*; D008113:Liver Neoplasms / Q000097:blood*; D015394:Molecular Structure; D011134:Polysaccharides / Q000097:blood / Q000737:chemistry; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000706:statistics & numerical data
D014408:Biomarkers, Tumor; C485359:HP protein, human; D006242:Haptoglobins; D011134:Polysaccharides; D005643:Fucose
Liquid chromatography−mass spectrometry; Meladrazine; cirrhosis; electrospray ionization; fucosylation; hepatocellular cancer