Comparison of heparin-binding to lactoferrin from human milk and from human granulocytes by means of affinity capillary electrophoresis.
Human lactoferrin from two different sources: milk and secondary granules of neutrophil granulocytes, was compared with respect to heparin binding by means of affinity capillary electrophoresis. This method is based on analysis of migration pattern shifts induced by various amounts of ligand present in the electrophoresis buffer. Since lactoferrin is a basic molecule, analyses were performed at pH 8 in a capillary with a neutral hydrophilic coating. However, analyzable peaks were only obtained in the presence of heparin in the electrophoresis buffer. Proportionally to the amount of heparin present, these peaks exhibited shape changes and strong migration shifts. In the ensuing analysis it could be demonstrated that the two lactoferrins had comparable migration shifts and peak shape changes. This indicates that also in this respect the two forms of lactoferrin are identical. Affinity capillary electrophoresis is a convenient and fast method to investigate functional characteristics of low amounts of unmodified biomolecules.
Heegaard
N H
NH
Department of Autoimmunology, Statens Serum Institute, Copenhagen, Denmark. nheegaard@ai.diag.ssi.dk
Brimnes
J
J
D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D007781:Lactoferrin / Q000737:chemistry / Q000378:metabolism*; D008895:Milk, Human / Q000737:chemistry*; D009504:Neutrophils / Q000737:chemistry*
D006493:Heparin; D007781:Lactoferrin