title: A MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) for total N-glycan analysis.
abstract: OBJECTIVE
  To develop a sensitive and quantitative method for monitoring the abnormal glycosylation of clinical and biopharmaceutical products.
  
  
  RESULTS
  MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) was proposed for sensitive and quantitative analysis of total N-glycans. The derivatization reactions (i.e., amidation of sialic acid and incorporation of a positive charge moiety into the reducing end) dramatically increased the linearity (R(2) > 0.99) and sensitivity (limit of detection is 0.5 pmol/glycoprotein) relative to underivatized glycans. In addition, the analytical strategy was chromatographic purification-free and non-laborious process accessible to the high-throughput analyses. We used MALDI-QTaG to analyze the N-glycans of α-fetoprotein (AFP) purified from normal cord blood and HCC cell line (Huh7 cells). The total percentages of core-fucosylated AFP N-glycans from Huh7 cells and normal cord blood were 98 and 18%, respectively.
  
  
  CONCLUSIONS
  This MALDI-MS-based glycomics technology has wide applications in many clinical and bioengineering fields requiring sensitive, quantitative and fast N-glycosylation validation.
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea.
authors.forename: Kyoung-Jin
authors.initials: KJ
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea.
authors.forename: Yoon-Woo
authors.initials: YW
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea.
authors.forename: Cheol-Hwan
authors.initials: CH
authors.lastname: Hwang
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea.
authors.forename: Han-Gyu
authors.initials: HG
authors.lastname: Park
authors.affiliation: Department of Microbial Engineering, College of Engineering, Konkuk University, Seoul, 143-701, Korea.
authors.forename: Yung-Hun
authors.initials: YH
authors.lastname: Yang
authors.affiliation: Immunoassay Tech. Lab, Healthcare Group, Future Technology R&D Division, SK Telecom, Anyang, Gyeonggi-do, 463-784, Korea.
authors.forename: Miyoung
authors.initials: M
authors.lastname: Koo
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea. ygkim@ssu.ac.kr.
authors.forename: Yun-Gon
authors.initials: YG
authors.lastname: Kim
mesh_terms: D002460:Cell Line; D005312:Fetal Blood; D054794:Glycomics / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry*; D022781:Hepatocytes; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D011134:Polysaccharides / Q000032:analysis*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D000509:alpha-Fetoproteins / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D000509:alpha-Fetoproteins
keywords: Abnormal glycosylation; Chemical derivatization; Glycosylation; MALDI-MS; N-Glycan; Quantitative analysis; UPLC