A high-throughput method for the quantification of iron saturation in lactoferrin preparations

作者：Majka Grzegorz; Spiewak Klaudyna; Kurpiewska Katarzyna; Heczko Piotr; Stochel Grazyna; Strus Magdalena; Brindell Malgorzata^*

来源：Analytical and Bioanalytical Chemistry, 2013, 405(15): 5191-5200.

DOI：10.1007/s00216-013-6943-9

• Lactoferrin
• Iron saturation
• Absorption ratio
• ICP-MS

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.

• 出版日期2013-6