A High-throughput Method for the Quantification of Iron Saturation in Lactoferrin Preparations

Overview

Journal Anal Bioanal Chem

Specialty Chemistry

Date 2013 Apr 23

PMID 23604471

Citations 28

Authors

Grzegorz Majka

Klaudyna Spiewak

Katarzyna Kurpiewska

Piotr Heczko

Grazyna Stochel

Magdalena Strus

Malgorzata Brindell

Affiliations

Soon will be listed here.

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.

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