Lactoferrin (formerly known as lactotransferrin) is a glycoprotein, and a member of a transferrin family, thus belonging to those proteins capable of binding and transferring Fe³⁺ ions. There are three forms of lactoferrin according to its iron saturation: apolactoferrin (iron free), monoferric form (one ferric iron), and hololactoferrin (binds two Fe³⁺). Lactoferrin is widely represented in various secretory fluids, such as milk, saliva, tears, and nasal secretions. Lactoferrin is one of the components of the immune system of the body; it has antimicrobial activity (bacteriocide, fungicide) and is part of the innate defence, mainly at mucoses. In particular, lactoferrin provides antibacterial activity to human infants. [1,2]

Bovine lactoferrin (Lf) supplementation has been implied in prevention of late-onset sepsis in preterm very low birth weight neonates [3]. However, little attention has been paid to iron saturation level of lactoferrin preparations used in reported studies. This seems alarming as there is evidence for Lf serving as iron donor for pathogen species. Biological impact of various lactoferrin species is still poorly understood and requires further research. Our goal is to differentiate effects mediated by apo- and hololactoferrin on both microorganisms and intestinal epithelial cells function. Also, we endeavour to obtain lactoferrin saturated with manganese ions (MnLf). It seems plausible that MnLf might tip the balance of intestinal microbial ecology in favour of probiotic species that require manganese for their growth.

Here, we present comprehensive method for quantification of metal saturation levels of bovine lactoferrin preparations based solely on the defined ratio of absorbances. Combining spectrophotometry, ELISA and ICP-MS we plotted calibration curve between metal saturation levels and absorbance ratio [4]. Also, several methods serving to obtain various lactoferrin species (MnLf included) were inspected and evaluated. Further studies included testing of antimicrobial activity and microbial adherence assays. These results will certainly contribute to better understanding of biological activities of lactoferrin species differentially saturated with metal ions in context of bacterial translocation from intestinal lumen to peripheral blood.

References

1  Adlerova L, Bartoskova A, Faldyna M. Lactoferrin: a review. Veterinarni Medicina, 53, 9, 457–468 (2008).

2   Levin RE, Kalidas S, Gopinadhan P, Pometto A. Food biotechnology. Boca Raton, FL: CRC/Taylor & Francis. p. 1028. ISBN 0-8247-5329-1 (2006).

3    Manzoni, P., Mostert, M. & Stronati, M. Lactoferrin for prevention of neonatal infections. Current Opinion in Infectious Diseases 24, 177–182 (2011).

4   Majka, G., Śpiewak K., Kurpiewska K., Strus M., Brindell M . A high-throughput method for the quantification of iron saturation in lactoferrin preparations. Anal Bioanal Chem 405, 5191–5200 (2013).

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