The advantage of modern agarose based chromatography sorbents for milk proteins separation and purification

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F16%3A43903439" target="_blank" >RIV/60461373:22330/16:43903439 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

The advantage of modern agarose based chromatography sorbents for milk proteins separation and purification

Original language description

Main criteria for choosing of chromatography media for milk proteins separation are cost of sorbents, binding capacity and work flow rate. For separation and purification of milk proteins previously are used sorbents with 200-500 ?m beads size. Over past five year has been made significant improvements of chromatography resins. The driver of this improvement was immunoglobulins, using of which is common in science and medicine. As result nowadays sorbents with binding capacity over 100 mg of immunoglobulins per ml of sorbent are present on the market. Some of these sorbents could be effectively used for milk proteins separation and purification. During our experiments were used two types of agarose cation exchange sorbent Bio-Works WorksBead 40S(Sweden) and Bio-Works WorksBead 100S (Sweden) Target proteins were ?-lactalbumin, lactoferrin and lactoperoxidase. For both sorbent dynamic binding capacity was over 70 mg/ml and was depended on feeding conditions and performance of column cleaning. Purification of ?-lactalbumin included several steps. The first one was precipitation technique using citric acid, centrifugation of precipitated apo ?-lactalbumin and final reconstitution of ?-lactalbumin by diluting in solution with calcium chloride. By using of this method, it was possible to get up to 75% pure ?-lactalbumin and main contaminant component was ?-lactoglobulin. For last step purification was used chromatography. In this case sorbent was used to bind ?-lactoglobulin, which has higher isoelectric point than ?-lactalbumin. For this separation was used acetic acid - sodium acetate buffer system without any gradient elution. The final purity of ?-lactalbumin after chromatographic purification was over 85%.

Czech name

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Czech description

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Type

O - Miscellaneous

CEP classification

GM - Food industry

OECD FORD branch

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Project

<a href="/en/project/QJ1210376" target="_blank" >QJ1210376: Colostrum as a source of new primary products in food and food supplements characterized by improved dietary properties and high content of natural biologically active substances.</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Publication year

2016

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů