Enrichment strategies are designed for the pretreatment
of low-abundance
glycans and glycopeptides prior to mass spectrometric (MS) analysis.
Here, a tip-based strategy is being reported for the enrichment of
glycopeptides and glycans using a piperazine modified polymeric monolithic
tip. The tip is fabricated using the free radical polymerization.
Fast separation (2 min) is achieved under optimized conditions with
20 cycles per step of loading, incubation, washing, and elution followed
by MALDI-MS analysis. A total of 25, 22, and 34 glycopeptides covering
all glycosylation sites are enriched by the modified tips from tryptic
digests of horse radish peroxidase, chicken avidin, and human immunoglobulin
G, respectively. Piperazine exhibits high selectivity 1:400 horse
radish peroxidase/bovine serum albumin, sensitivity to 100 attomoles,
recovery 89.51%, and batch to batch reproducibility (RSD > 1) in
glycopeptides
enrichment. Piperazine tips also enrich glycans from ovalbumin and
human immunoglobulin G. High selectivity (1:1200, ovalbumin/BSA) and
detection limit of 100 attomole is attained for glycans and furthermore
58 glycans are enriched from human serum. Thus, piperazine tips can
be used as an enrichment tool for swift, cost-effective routine analysis
of biological samples for separation of glycopeptides and glycans.
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