N-glycome analysis of dried blood spots from different blood preparations and its potential for pre-diabetes and diabetes distinction

Dried blood spot (DBS) sampling has gained popularity as a more convenient and safer alternative to traditional blood sampling methods. One promising application of DBS analysis is the examination of N-glycosylation, a post-translational modification of proteins linked to various biological and pathological functions. In this study, researchers employed a protocol involving ultra-high-performance liquid chromatography based on hydrophilic interactions and fluorescence detection (HILIC-UHPLC-FLR) to profile N-glycosylation in DBS.

The protocol includes cutting the DBS, extracting the proteins, enzymatic digestion, labeling with 2-aminobenzamide, cleanup, and measurement using HILIC-UHPLC-FLR.

The researchers compared DBS N-glycosylation profiles with those of plasma and discovered that DBS maintain a stable N-glycosylation profile regardless of whether they are collected from fresh blood, frozen whole blood, or a combination of frozen blood cells and corresponding frozen plasma. Furthermore, the researchers examined DBS N-glycans from individuals with pre-diabetes and diabetes and found that fucosylation, bisection, and galactosylation showed a statistically non-significant increasing trend in diabetes, while sialylation demonstrated a statistically non-significant decreasing trend in diabetes.

The primary advantage of this method is its ability to repurpose samples initially intended for other purposes, such as frozen whole blood. Moreover, DBS N-glycan profiling offers a simpler, more cost-effective, and less invasive approach to diagnosing and monitoring pre-diabetes and diabetes patients compared to conventional plasma analysis.