Fetuin, bovine

Cell Culture-Derived Media-Derived Protein Impurities. Immunoassays can detect low impurity levels (<1 ppm).4 The ELISA is probably one of the most sensitive analytical methods. If bovine serum is used as a media component, then testing should include ELISAs for bovine serum albumin (BSA), bovine transferrin, bovine fetuin, and bovine IgG. Often hormones and growth factors, such as insulin or insulinlike growth factor, are used as media components. ELISAs should be used to detect and quantitate these residuals in the various production steps as well as in the final product. There are commercially available antibodies to most commonly used media components. If proprietary media components are used, then the same investment in time and effort is required for the production of specific antibodies, as described above for host cell impurities. 

The strategy used to perform the PMP oligosaccharide profile analysis, with the use of bovine fetuin as a model system, is given in Scheme 1. Bovine fetuin, a major glycoprotein in fetal calf serum, has been widely used as a model for the study of glycoprotein structure. This glycoprotein contains both N- and O-linked oligosaccharides and detailed structural information is available (9-11). 

With bovine fetuin as a model system, the use of PMP labeling in the routine rp-HPLC profile analysis of glycoproteins has been evaluated. Comparable profiles for oligosaccharides can be obtained regardless of whether they are released by automated chemical or manual enzymatic treatments. The PMP-oligosaccharide samples recovered fi om HPLC are suitable for MALDI TOP and LC ESI mass spectrometric analysis. The presence of all three N-linked and one of the 0-linked oligosaccharides was confirmed. 

SCHEME 15.20 Release of a tri-antennary complex type oligosaccharide from bovine fetuin allowed preparation of asparagine building block 172. 

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... 

Bovine Fetuin. See Glycoproteins Bovine Serum Albumin Sol n.. See Serum albumin... 

Uses Skin conditioner, hair conditioner in cosmetics, hair care, skin care Trade Name Synonyms Bovine Fetuin t[lntergen http //www.intergenco.com], Glyprosol 20 [Arch Personal Care Prods. http //www.archchemicals.com], Glyprosol ... 

Saw palmetto (Serenoa serrulata) extract 84604-16-0 Bovine Fetuin Glycoproteins Glyprosol 20 Glyprosol SD Immucell ... 

FIGURE 19.7 Comparison between protein ISTD and stable-isotope-labeled peptide ISTD (a) Use of bovine fetuin as ISTD for the mAbX assay. The r2 of the linear regression was 0.9943. (b) Use of stable-isotope-labeled peptide as ISTD for the mAbX assay. The r2 of the linear regression was 0.9886. [Reprinted from Yang et al. (2007) with permission of the American Chemical Society.]... 

FIGURE 4 HPLC chromatograms of the peptides produced from bovine fetuin by trypsin and Asp-N as measured by the electrospray mass spectrometer (A) Total ion current (TIC) (B) reconstructed ion chromatogram (RIC) for ion 204. Glycopeptides containing residues Asn 81, Asn 138, Asn 158, Ser 253, and Thr 262/Ser 264 were observed in the LC/MS analysis. Reprinted, with permission, from Carr et al. (1993). 

Although amino acids and peptides can also be electrochemically detected under the conditions being used [218], Hardy et al. [261] could not identify any interfering signals in the monosaccharide retention range when analyzing hydrolysates of bovine fetuin and fibrinogen. UV active components of these protein hydrolysates elute either in the column void volume or far behind the monosaccharides. 

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

Sampies (top trace) 2AB-labeled glucose homopolymer (GHP) ladder, which is used as an external standard (middle trace) 2AB-labeled N-glycans released from human IgG (bottom trace) 2AB-labeled N-glycans released from bovine fetuin. Retention times of glycans shown as peak labels in the chromatograms and are expressed in terms of glucose units (GU) by reference to the GHP ladder. 

Samples (top trace) 2AA-labeled monosaccharide standards (bottom trace) 2AA-labeled monosaccharides released from bovine fetuin glycoprotein. 

The profiles for the analysis of the neutral and acidic monosaccharides released from bovine fetuin are shown in Figures 8.9 and 8.10. The quantitative results (Table 8.1) are in line with published data for a multicenter study [18]. 

TABLE 8.1 REFERENCES Quantification of Monosaccharides in 201 Bovine Fetuin (nmoles/mg protein) ... 

---