Boronic acid functionalized fibrous cellulose for the selective enrichment of glycopeptides

• Published in: Journal of separation science 2020-04, Vol.43 (7), p.1348-1355
• Main Authors: Sajid, Muhammad Salman, Jabeen, Fahmida, Hussain, Dilshad, Gardner, Qurra‐tul‐Ann Afza, Ashiq, Muhammad Naeem, Najam‐ul‐Haq, Muhammad
• Format: Article
• Language: English
• Subjects: Acids; Adsorption; Animals; Antibiotics; boronic acid; Boronic Acids - blood; Boronic Acids - chemistry; Boronic Acids - metabolism; Cattle; Cellulose; Cellulose - blood; Cellulose - chemistry; Cellulose - metabolism; Cellulose fibers; Enrichment; Fourier transforms; Glycopeptides; Glycopeptides - blood; Glycopeptides - chemistry; Glycopeptides - metabolism; glycopeptides enrichment; Glycoproteins; Horseradish Peroxidase - metabolism; human serum; Humans; Immunoglobulins - metabolism; mass spectrometry; Morphology; Peroxidase; Polysaccharides; Proteins; Serum albumin; Serum Albumin, Bovine - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.201900983

Enrichment of glycoproteins has been important because of their dynamicity and role in biological systems. Study of glycoproteins is complex because of the simultaneous glycosylation and deglycosylation inside the body. Often employed affinities for glycopeptides are hydrazide, boronic acid, or physiosorbed lectin on support materials. Cellulose, a natural polysaccharide, has rich surface chemistry, stable structure, low cost and availability in different variants. In present study, fibrous cellulose is oxidized using periodate to modify with boronic acid. Attachment of boronic acid is confirmed by Fourier transform infrared spectroscopy. Particle size and morphology of boronic acid@fibrous cellulose is studied by scanning electron microscopy. The enrichment efficiency is evaluated by using horseradish peroxidase as model protein. Boronic acid@fibrous cellulose is selective up to 1:250 for spiked horseradish peroxidase in bovine serum albumin digest, sensitive down to 0.1 femtomol and recovering 88.15% glycopeptides. Moreover, protein binding capacity is determined as 213 mg/g and 41% sequence coverage of horseradish peroxidase protein with all eight glycosylation sites detected. Total of 18 glycopeptides are enriched from immunoglobulin digest showing ability of boronic acid@fibrous cellulose to enrich glycoproteins from multiglycoforms. Enrichment from human serum recovers 18% extracellular and 72% secreted glycoproteins via bottom‐up approach and online tools.