Brush polymer modified and lectin immobilized core-shell microparticle for highly efficient glycoprotein/glycopeptide enrichment.
Protein glycosylation regulates numerous important biological processes and plays key roles in many diseases including cancer, diabetes and inflammation. The ability to efficiently profile variation of protein glycosylation in biological samples is very useful for identifying new diagnostic biomarkers or developing new therapeutic approaches. Due to the low availability of glycoprotein/glycopeptide from natural sources, enrichment before mass spectrometry (MS) analysis is usually a prerequisite. Affinity enrichment using lectins is currently one of the most widely adopted approaches. Conventionally, lectins are immobilized on solid supporting materials for sample recovery. However, the limited specific surface area, high steric hindrance and rigid nature of such supporting materials restricts lectin loading amount and results in low flexibility as well as accessibility of the immobilized lectins. Therefore, we proposed using core-shell microparticles composed of silica core and brush-like polymer chains shell for improved lectin immobilization. The surface bound brush-like polymer are synthesized by in situ growth of polymer chains from microparticle surface using surface initiated atom transfer radical polymerization (SI-ATRP). The flexible non-crosslinked polymer chains not only provide numerous binding sites, but also work as three-dimensional support for lectin immobilization, which leads to high loading amount and good accessibility of the immobilized lectin. Successful enrichment which facilitated glycoprotein/glycopeptide identification is demonstrated.
School of Life Science and Technology, Beijing Institute of Technology, Beijing 100081, PR China; National Center for Protein Sciences Beijing, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, PR China.
Yiting
Y
Pan
Haihong
H
Bai
Cheng
C
Ma
Yulin
Y
Deng
Weijie
W
Qin
Xiaohong
X
Qian
D000595:Amino Acid Sequence; D003208:Concanavalin A / Q000737:chemistry*; D006020:Glycopeptides / Q000302:isolation & purification*; D006023:Glycoproteins / Q000302:isolation & purification*; D006031:Glycosylation; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D008768:Methylmethacrylates / Q000737:chemistry; D008969:Molecular Sequence Data; D058105:Polymerization; D012822:Silicon Dioxide; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014909:Wheat Germ Agglutinins / Q000737:chemistry*
D006020:Glycopeptides; D006023:Glycoproteins; D055767:Immobilized Proteins; D008768:Methylmethacrylates; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D012822:Silicon Dioxide
Enrichment; Glycoprotein/glycopeptides; Lectin; SI-ATRP