Simple purification of immunoglobulins from whey proteins concentrate.
We have developed a new protocol with only two steps for purification of immunoglobulins (Ig) from a protein concentrate of whey. Following this protocol, we have an 80% recovery of immunoglobulins, fairly pure. The purification was achieved by eliminating the BSA, via a strong adsorption on DEAE-agarose. Full desoprtion of the other serum proteins could be achieved without contamination with BSA. Thus, a protein solution containing only Ig and very small proteins (e.g., beta-lactoglobulins and alpha-lactalbumin) was obtained. Offering this protein mixture to a lowly activated aminated support, only Ig adsorbed on the support. It has been shown that BSA is able to interact with other proteins (including Ig and lactalbumins). This ability to form complexes with other proteins prevented the success of the direct adsorption of Ig on this mildly activated support, even although Ig should be the largest protein presented in dairy whey.
Pessela
Benevides C C
BC
Departamento de Biocatalisis, Instituto de Catalisis-CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain.
Torres
Rodrigo
R
Batalla
Pilar
P
Fuentes
Manuel
M
Mateo
Cesar
C
Fernández-Lafuente
Roberto
R
Guisán
José M
JM
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005021:Ethylamines; D007136:Immunoglobulins / Q000302:isolation & purification* / Q000378:metabolism; D008894:Milk Proteins / Q000378:metabolism; D012685:Sepharose / Q000031:analogs & derivatives; D000067816:Whey Proteins
D005021:Ethylamines; D007136:Immunoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins; C083051:monoaminoethyl-N-aminoethyl-agarose; D012685:Sepharose