engleski

Application of protein hydrolysates from flaxseed and hempseed oil cakes in animal cell culture media formulation

A lot of industries are looking for efficient exploitation of organic residues from various domains of agriculture and food manufacturing. Oil cake, a substance that remains after oil extraction from oil plant seeds, is one of these organic residues with great potential of application. Because of their protein content, the flaxseed and hempseed oil cakes can potentially be used in animal cell technology. Animal cells are complex expression system applied in wide range of biotech and biomedical techniques. Cell growth supporting microenvironment is largely defined by nutrient composition of culture media. Content of the proteins as much as their origin have a key role in culture media preparation. Fetal bovine serum (FBS) is still used as a major protein source, despite its compositional inconsistency and safety related risks. To overcome these problems, cell culture experts developed media where serum proteins can be partially or fully replaced with non-animal origin proteins. In this work we used plant protein hydrolysates derived from flaxseed and hempseed oil cakes. They both contain about 20% proteins, and as such can be processed into nutrient media supplement for animal cell cultivation. The main objective of this study is to determine effects of flaxseed and hempseed oil cake protein hydrolysates on growth and productivity of recombinant protein producing CHO cell line. To obtain the hydrolysates, proteins were isolated from hempseed and flaxseed oil cake by extraction and precipitation steps. The hydrolysis of protein isolates was carried out with commercial microbial proteases. CHO cell cultivation was performed in standard growth media containing FBS partially replaced with total protein hydrolysates. Protein content of the culture media was determined using SDS-PAGE and colorimetric techniques. CHO cell growth was monitored with hemocytometer and recombinant protein production by immunodetection. Our preliminary results indicate that oil cake protein hydrolysates show potential of being a beneficial culture media supplement. Both hempseed and flaxseed protein hydrolysates have positive effects on cell growth and volumetric production when compared to medium without hydrolysates added.

flaxseed ; hempseed ; oil cakes ; protein hydrolysates ; animal cells

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