A high-throughput method for measurement of glycohemoglobin in blood samples utilizing laser-accelerated proteolysis and MALDI-TOF MS

作者：Li, Lanting; Zang, Wenjuan; Zhang, Xiangmin^*

来源：Analytical and Bioanalytical Chemistry, 2016, 408(5): 1507-1513.

DOI：10.1007/s00216-015-9258-1

• N-termini
• HbA1c
• Post-translational modification
• Laser-accelerated proteolysis

Glycosylated hemoglobin A1c (HbA1c) is a useful marker for the diagnosis of diabetes mellitus. Commercial column separation methods for HbA1c measurement were lacking throughput and sometimes interfered with hemoglobin variants. In this work, we developed a high-throughput and specific method for HbA1c by quantitative measurement of N-terminal peptides (NT method). Two thousand specimens could be measured in 8 h. The high-throughput was achieved by using a fast analysis of matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and an efficient proteolysis accelerated by laser irradiation. An intensity ratio of glycosylated to non-glycosylated hemoglobin N-terminal peptides was used to calculate the HbA1c level in blood. Interference from Hb variants of N-terminal peptides could be excluded by a highly accurate mass selection. The coefficient of variation (CV) of intra-assay precision was 9.8 and 9.9 %, respectively. The CVs of inter-assay precision over 20 days were 9.1 and 8.4 %, respectively. Measurement results were well correlated with the commercially available column method (r = 0.995). The NT method is promising for large-scale screening for diabetes mellitus among people.

• 出版日期2016-2
• 单位复旦大学