TY - JOUR
T1 - Direct Screening of Glycan Patterns from Human Sera
T2 - A Selective Glycoprotein Microarray Strategy
AU - Tu, Hsiu Chung
AU - Lee, Yen Pin
AU - Liu, Xuan Yu
AU - Chang, Chuan Fa
AU - Lin, Po Chiao
N1 - Funding Information:
This work is supported by grants from Ministry of Science and Technology (MOST103-2113M-110-010 and MOST 104-2627M-007-003), National Sun Yat-sen University, and Kaohsiung Medical University Center for Research Resources and Development (NSYSU-KMU JOINT RESEARCH PROJECT, #NSYSUKMU 107-PO15). The Typhoon 9410 imaging system was support by Center for Research Resources and Development of Kaohsiung Medical University.
Publisher Copyright:
Copyright © 2019 American Chemical Society.
PY - 2019/3/18
Y1 - 2019/3/18
N2 - Protein glycosylation is one of the most complicated but significant post-translational modifications. Minor alterations in glycan structure can considerably affect the biology of a cell. Therefore, direct monitoring of glycan patterns of glycoproteins is closely related to cancer progression as well as metastasis. In this study, a boronic acid (BA)-tosyl-directed strategy to selectively immobilize glycoproteins on glass slides was successfully developed even in the presence of high-abundant nonglycosylated proteins. To enhance the immobilization efficiency and reduce the undesired nonspecific absorption, the strain-promoted alkyne azide cycloaddition (SPAAC) conjugation chemistry and surface blocking conditions were carefully optimized for the collection of reliable data. The optimized glycoprotein microarray platform describes specific lectin-recognition patterns of glycoproteins of interest in E. coil lysate and fetal bovine serum (FBS), which encourages us for direct monitoring of glycan patterns from human sera without tedious sample preparation. Three serum groups comprised of healthy controls and lung cancer and pancreatic cancer patients were analyzed by this new technique. Remarkably, the distinguishable glycan patterns of the three groups make them a powerful platform for cancer screening and prediagnosis.
AB - Protein glycosylation is one of the most complicated but significant post-translational modifications. Minor alterations in glycan structure can considerably affect the biology of a cell. Therefore, direct monitoring of glycan patterns of glycoproteins is closely related to cancer progression as well as metastasis. In this study, a boronic acid (BA)-tosyl-directed strategy to selectively immobilize glycoproteins on glass slides was successfully developed even in the presence of high-abundant nonglycosylated proteins. To enhance the immobilization efficiency and reduce the undesired nonspecific absorption, the strain-promoted alkyne azide cycloaddition (SPAAC) conjugation chemistry and surface blocking conditions were carefully optimized for the collection of reliable data. The optimized glycoprotein microarray platform describes specific lectin-recognition patterns of glycoproteins of interest in E. coil lysate and fetal bovine serum (FBS), which encourages us for direct monitoring of glycan patterns from human sera without tedious sample preparation. Three serum groups comprised of healthy controls and lung cancer and pancreatic cancer patients were analyzed by this new technique. Remarkably, the distinguishable glycan patterns of the three groups make them a powerful platform for cancer screening and prediagnosis.
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U2 - 10.1021/acsabm.9b00001
DO - 10.1021/acsabm.9b00001
M3 - Article
C2 - 35021376
AN - SCOPUS:85072846939
SN - 2576-6422
VL - 2
SP - 1286
EP - 1297
JO - ACS Applied Bio Materials
JF - ACS Applied Bio Materials
IS - 3
ER -
