Fractioning of Salmon Protein Hydrolyzate

For the past one year, Hofseth BioCare has been pursuing a research project together with students at St. Xavier's College to isolate the active components present in our salmon protein hydrolysate (ProGo™) that may be responsible for the excellent increase in iron uptake and hemoglobin in anaemic humans.

ProGo™ is a protein hydrolysate that contains approximately 627 individual peptides (MALDI-TOF analysis) ranging from dimers (containing 2 amino acids) to 25-mer (containing 25 amino acids). The initial aim of the research project was to investigate if the iron uptake bioactivity could be focused into a more narrow range of peptides. 

The hydrolysate powder was fractionated into approximately 500 Dalton size fragments using a size occlusion column methodology developed by HBC.  This lead to 5 fractions of peptides - from 110-1000 Dalton; 1000-1500 Dalton; 1500-2000 Dalton; 2000-2500 Dalton and > 2500 Dalton.
These five fractions were compared to the total hydrolysate powder for iron uptake increase activity using the CACO-2 in-vitro cell method developed by us earlier.

When the results were normalized to 100% iron uptake for ProGo™ powder, the 5 fraction showed the following bioactivity:

110-1000 Dalton =  <10%
1000-1500 Dalton = <10%
1500-2000 Dalton = 77%
2000-2500 Dalton = 30%
2500 Dalton = < 10%

This indicates that the iron uptake increase bioactivity is maximally preserved in the 1500-2000 Dalton fraction.  HPLC analysis of the this fraction showed approximately 48 distinct peptides of unknown structure.  Thus, the size occlusion chromatography has led to at least a 12X concentration of iron uptake bioactivity. (627 peptides gave 100% activity vs 48 peptides have shown 77% activity)
We are in the process of trying to further concentrate the bioactivity using Gel Fractionation on the basis of charge/polarity.  We are also trying to arrange for LC-MS analysis to identify the peptides that will be the most active compounds after concluding the concentration work.
Hofseth BioCare ASA aim to develop a future Rx product, based on the findings, for treatment of anemia.