Naturally occurring anti-band 3 antibodies bind to protein rather than to carbohydrate on band 3

Laboratory for Biochemistry, Swiss Federal Institute of Technology, ETH-Zentrum, Zurich.
Journal of Biological Chemistry (Impact Factor: 4.57). 12/1993; 268(31):23562-6.
Source: PubMed


Naturally occurring anti-band 3 antibodies were affinity purified from pooled human IgG (Sandoglobulin) (Lutz, H. U., Flepp, R., and Stringaro-Wipf, G. (1984) J. Immunol. 133, 2610-2618). They bound to the major integral membrane protein of human red blood cells and its 55-kDa NH2-terminal chymotryptic fragment but not to the carbohydrate-rich 38-kDa fragment on blots. Likewise, neither an endo-beta-galactosidase nor a neuraminidase treatment of band 3 on intact red cells reduced their binding to the blotted antigen. Lactoferrin (10 micrograms/ml) had no significant effect on their binding to band 3 and to its 55-kDa chymotryptic fragment. Even in the presence of 20 micrograms/ml lactoferrin anti-band 3 antibodies bound specifically to chymotrypsin-pretreated and oxidatively stressed red cells. Thus, naturally occurring anti-band 3 antibodies bind to protein rather than carbohydrate within band 3 protein, irrespectively of whether the antibodies were depleted of anti-idiotypic and other IgG-reactive antibodies or not.


Available from: Hans Ulrich Lutz
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    • "A detailed analysis further showed that binding of anti-band 3 NAbs to blotted band 3 protein from RBC membranes was neither inhibited by pretreating RBC with neuraminidase nor endo-β-galactosidase. In addition, its binding to band 3 and to the 55 kDa fragment of band 3 was not inhibited at all by 10 μg/ml lactoferrin (Lutz et al., 1993a). This was in complete contrast to the properties of Beppu's “anti-band 3 antibodies,” for which these authors showed an 80% inhibition of binding to oxidatively stressed RBC by 10 μg/ml lactoferrin (Beppu et al., 1992). "
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