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: Grzegorz Majka, Klaudyna Śpiewak, Katarzyna Kurpiewska, Piotr Heczko, Grażyna Stochel, Magdalena Strus, Małgorzata Brindell
• Journal: Analytical and bioanalytical chemistry (Anal Bioanal Chem) Vol. 405 Issue 15 Pg. 5191-200 (Jun 2013) ISSN: 1618-2650 [Electronic] Germany
• PMID: 23604471 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Apoproteins
• apolactoferrin
• Iron
• Lactoferrin

Topics

• Apoproteins (chemistry)
• Chromatography (methods)
• Iron (chemistry)
• Lactoferrin (chemistry)