Instructions for Use - Glyco Kit MB-LAC AIA - Bruker

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Figure 1 exemplarily depictured possible experimental strategies to facilitate the identification of glyco-proteins, to enable the detection of N-glycosylation sites, to support the structural characterization of N-glycosylated peptides and to perform glyco-profiling studies in the clinical context. General Information Glycosylation is the most common post-translational modification of proteins, playing a key role in protein folding, stability, solubility, activity and molecular recognition. Altered glycosylation is known to act as an indicator or effector in pathologic mechanisms with potential impact on diagnosis and therapy of human diseases. Immunity to certain infectious diseases, prion diseases and cancer can all be associated with abnormal glycosylation patterns. As an example, increased branching and sialylation of N-linked glycans on cancer cells are potential tumor markers in molecular diagnostics. Bruker Daltonics’ CLINPROT magnetic glyco-beads, specifically functionalized with lectins or boronic acids that bind to various structural motifs, facilitate the selective isolation of different glycoproteins out of complex biological sources (Fig. 1). Bruker Daltonics has developed workflows combining the specific pre-fractionation employing glyco-specific magnetic particles, e.g. ConA, WGA, LCA or boronic acids coated beads, and LC-MALDI analysis to identify glycoproteins, to locate N-glycosylation sites and to get structural information about N-glycosylated peptides. MB-LAC Con A MB-LAC LCA MB-LAC WGA MB-LAC AIA MB-CovAC boronic High-mannose type Hybrid type Complex type Glycorotein Glycoprotein Glycoprotein Man GlcNAc Gal Fuc NeuAc GalNAc Glycoprotein Glycoprotein Glycoprotein O-linked glycans C-linked glycans N-linked glycans Fig. 2: Binding motifs of Bruker Daltonics’ CLINPROT TM magnetic glyco-beads. The different beads represent overlapping but individual binding profiles. Successful applications and reproducibility of Bruker Daltonics’ CLINPROT TM magnetic glycobeads using model systems or complex samples like serum, plasma or urine were demonstrated in various studies 1,2,3 . Further, long term stability and storage conditions were extensively investigated to assure the high quality of Bruker Daltonics’ CLINPROT TM magnetic glycobeads 4 .The application of glyco-beads in the context of glyco-profiling and biomarker discovery was closely described by Drake et al. 5 Page 2 of 19
Table of Contents General Information........................................................................................................................... 2 Intended Use..................................................................................................................................... 4 Product description............................................................................................................................ 4 Kit Components................................................................................................................................. 5 Storage and lifetime information........................................................................................................ 5 Risk and safety information ............................................................................................................... 5 General remarks for sample handling and MALDI measurements ................................................... 6 Recommended sample preparation procedure ................................................................................. 7 Comments ......................................................................................................................................... 9 MALDI-TOF MS Target Preparation................................................................................................10 Cleaning of MALDI-MS targets.................................................................................................... 10 Cleaning of AnchorChip TM targets ........................................................................................... 10 Cleaning of polished steel targets ........................................................................................... 11 Matrix preparation on the MALDI-MS targets.............................................................................. 12 Analysis of glycoproteins using 2,5-DHAP .............................................................................. 12 Analysis of glycopeptides using 2,5-DHB................................................................................ 13 Analysis of deglycosylated peptides using PAC targets.......................................................... 15 Example .......................................................................................................................................... 16 Isolation of Asialofetuin from a protein mixture ........................................................................... 16 References...................................................................................................................................... 17 Trouble shooting.............................................................................................................................. 18 Ordering Information ....................................................................................................................... 19 Support............................................................................................................................................ 19 Page 3 of 19
Page 1: CARE products are designed for supp
Page 5 and 6: Kit Components Materials provided i
Page 7 and 8: Recommended sample preparation proc
Page 9 and 10: 19. Place the tube on the Magnetic
Page 11 and 12: Cleaning of polished steel targets
Page 13 and 14: 8. 1 µl of the turbid suspension h
Page 15 and 16: Analysis of deglycosylated peptides
Page 17 and 18: References 1 Sparbier K, Koch S, Ke
Page 19: Ordering Information Product Size (

Instructions for Use - Glyco Kit MB-LAC AIA - Bruker

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