title: A high-throughput method for measurement of glycohemoglobin in blood samples utilizing laser-accelerated proteolysis and MALDI-TOF MS.
abstract: 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.
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Lanting
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Zang
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200433, China. xmzhang@fudan.edu.cn.
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Zhang
mesh_terms: D015415:Biomarkers / Q000097:blood*; D001800:Blood Specimen Collection / Q000379:methods; D006442:Glycated Hemoglobin / Q000032:analysis*; D057166:High-Throughput Screening Assays; D006801:Humans; D007834:Lasers; D059748:Proteolysis / Q000528:radiation effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D015415:Biomarkers; D006442:Glycated Hemoglobin; C517652:hemoglobin A1c protein, human
keywords: HbA1c; Laser-accelerated proteolysis; N-termini; Post-translational modification