Fetuin separation

Fig. 3-120. Separation of two structurally similar undecasaccharides from asialo fetuin. — Separator column CarboPac PA-1 eluent (A) 0.15 mol/L NaOH, (B) 0.15 mol/L NaOH + 1 mol/L NaOAc gradient linear, 50% B to 100% B in 20 min detection and injection volume see Fig. 3-105.

The separation of ANTS-derivatized glycans released from bovine RNase B, bovine fetuin, and chicken ovalbumin was reported by means of ion-pair RPLC with trimethylammonium as ion-pairing agent [79]. 

There are a few examples where mixtures of complex oligosaccharide have been isolated from natural sources such as bovine fetuin, porcine fibrinogen or conalbumin, and the mixture converted to the glycosyl amine and then coupled to an amino acid before separation of the individual components by HPLC. In one example described, 15 N-linked... 

Recently, a new polyacrylamide (PAA) derivative that allows dynamic coating (UltraTrol LN- Target Discovery, Palo Alto, CA) has been reported for the efficient separation of glycoforms of ATT [70], A comparison of the ability of PB and UltraTrol LN for the separation of bovine alpha-1-acid glycoprotein (AGP), bovine serum fetuin (BSF), and rhEPO sialoforms and their application coupling with MS has been reported [30,71]. Better resolution of sialoforms was obtained with capillaries coated with UltraTrol-LN in acidic volatile buffers used for coupling CE to ESI-TOF MS. 

The importance of additives to the separation buffer, even for coated capillaries, should be emphasized here. Both zwitterionic buffers and tensioactive additives have been shown to improve the separation and recovery of rhtPA with PVA-coated capillaries [77]. On DBl capillaries, a dimethylpolysiloxane coating widely used in gas chromatography (GC), optimal separations of fetuin, rhEPO, and AGP isoforms were obtained with acidic acetate buffers containing 0.4-0.5% (w/v) hydroxypropylmethycellulose (HPMC). In this case, the addition of a neutral polymer to the separation buffer makes the EOF almost negligible and protein interactions with the inner surface of the capillary were prevented [78]. 

Figure 8.84 Separation of native N-glycans from bovine fetuin with either charged aerosol or MS detection. Coiumn GlycanPac AXH-1,...

FIGURE 8.4 Separation of fetuin N-glycans by WAX, followed by HILIC. WAX... 

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