title: A high-throughput method for the quantification of iron saturation in lactoferrin preparations.
abstract: Lactoferrin is considered as a part of the innate immune system that plays a crucial role in preventing bacterial growth, mostly via an iron sequestration mechanism. Recent data show that bovine lactoferrin prevents late-onset sepsis in preterm very low birth weight neonates by serving as an iron chelator for some bacterial strains; thus, it is very important to control the iron saturation level during diet supplementation. An accurate estimation of lactoferrin iron saturation is essential not only because of its clinical applications but also for a wide range of biochemical experiments. A comprehensive method for the quantification of iron saturation in lactoferrin preparations was developed to obtain a calibration curve enabling the determination of iron saturation levels relying exclusively on the defined ratio of absorbances at 280 and 466 nm (A(280/466)). To achieve this goal, selected techniques such as spectrophotometry, ELISA, and ICP-MS were combined. The ability to obtain samples of lactoferrin with determination of its iron content in a simple and fast way has been proven to be very useful. Furthermore, a similar approach could easily be implemented to facilitate the determination of iron saturation level for other metalloproteins in which metal binding results in the appearance of a distinct band in the visible part of the spectrum.
authors.affiliation: Department of Inorganic Chemistry, Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow, Poland.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Majka
authors.forename: Klaudyna
authors.initials: K
authors.lastname: Śpiewak
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kurpiewska
authors.forename: Piotr
authors.initials: P
authors.lastname: Heczko
authors.forename: Grażyna
authors.initials: G
authors.lastname: Stochel
authors.forename: Magdalena
authors.initials: M
authors.lastname: Strus
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Brindell
mesh_terms: D001059:Apoproteins / Q000737:chemistry; D002845:Chromatography / Q000379:methods; D007501:Iron / Q000737:chemistry*; D007781:Lactoferrin / Q000737:chemistry*
chemical_list: D001059:Apoproteins; C039223:apolactoferrin; D007501:Iron; D007781:Lactoferrin