Fucose antibodies

The anti-fucose antibodies were eluted by 0.5 M L-fucose solution from the first column and the anti-BSA antibodies by elution with 0.5 M ammonium thiocyanate from the second. Agar diffusions are shown in the plates in Fig. 14. 

On gel electrophoresis, the purified antibody preparations yield single bands when the gel is stained for proteins. However, on gel isoelectrofocus-ing, differences in the results maybe noted, as shown in Fig. 15. Several protein isomers were present in each antibody preparation, with 7 isomers being detected in the anti-BSA antibodies (B) and 11 isomers in the anti-fucose antibodies (F). The coupled electrofocusing-agar diffusion method showed that each isomer of the anti-fucose set possessed the same antibody activity with the antigen, a-L-fucosyl-BSA. 

Fucose was tested as an inhibitor for both types of antibodies by the micro-inhibition test. These results are presented in Fig. 16. The center well of plate A contains native anti-fucose antibodies, with the native antibodies diffusing against decreasing concentrations (20 to 1 jug) of Fuc-BSA in the outer wells (1 to 6). The anti-fucose antibody yielded precipitin bands with 4 concentrations of antigen. Antibodies incubated with L-fucose (plate B) yielded a precipitin complex only at the highest concentration of antigen. A calculation from the concentrations shows that the L-fucose... 

Fig. 14.—Agar-diffusion plates of anti-fucose antibodies and anti-BSA antibodies. F = fucose-BSA, B = BSA. xf, xB = periodate-oxidized samples, Se = anti-BSA antibodies, and A = anti-fucose antibodies. (Reprinted with permission from Journal of Protein Chemistry, Volume 13, J. H. Pazur, B. Liu, and T. F. Witham, pp. 59-66, copyright 1994 Journal of Protein Chemistry.)...

Fig. 16.—Inhibition of anti-Fuc and anti-BSA antibodies by fucose. A, = anti-fucose antibodies, I = anti-fucose antibodies + fucose, A2 = BSA antibodies, L = BSA antibodies + fucose.

Wilson, I.B.H., Harthill, J.E., Mullin, N.P., Ashford, D.A. and Altmann, F. (1998) Core al,3-fucose is a key part of the epitope recognized by antibodies reacting against plant JV-linked oligosaccharides and is present in a wide variety of plant extracts. Glycobiology 8, 651-661. 

C5-1 antibody was produced in alfalfa, the glycan component consisted predominantly of a mature oligosaccharide comprising a core a(l,3)-fucose residue, a bisecting P(l,2)-xylose residue and two terminal GlcNAc residues (Fig. 15.4) [6],... 

One of the applications of CE for carbohydrate analysis that has shown increase acceptance in the biotechnology industry is the relative distribution of the N-linked oligossacharides on the conserved asparagines residue on the Fc portion of rMAbs. The advantages of CE over traditional methods include enhanced separation efficiencies, shorter analysis time, and improved robustness. The oligosaccharides present on a monoclonal antibody are asialo-N-linked complex biantennary structures with a core fucose, typical of the structures produced in the Chinese hamster ovary (CHO) cells. The main forms share the fucosylated branched core structure but vary in their terminal galactose occupancy ... 

Shields RL, Lai J, Keck R et al. Lack of fucose on human IgGlN-linked oligosaccharide improves binding to human Fcgamma RIII and antibody-dependent cellular toxicity. J Biol Chem 2002 277 26733-26740. 

Niwa R, Hatanaka S, Shoji-Hosaka E et al. Enhancement of the antibody-dependent cellular cytotoxicity of low-fucose IgGl Is independent of FcgammaRIIIa functional polymorphism. Clin Cancer Res 2004 10 6248-6255. 

Shinkawa T, Nakamura K, Yamane N, shoji-Hosaka E, Kanda Y, Sakurada M, Uchida K, Anazawa H, Satoh M, Yamasaki M, Hanai N, Shitara K. The absence of fucose but not the presence of galactose or Bisecting A-Acetylglucosamine of human IgGl complex-type oligosaccharides shows the critical role of enhancing the antibody-dependent cellular cytotoxicity. J Biol Chem 2003 278 3466-3473. 

Antibodies Carbohydrates Galactose Mannose Fucose Xylosyl Folic acid Transferrin... 

---