title: Affinity chromatography matrices for depletion and purification of casein glycomacropeptide from bovine whey.
abstract: Casein glycomacropeptide (CMP) is a 64- amino acid peptide found in cheese whey, which is released after κ-casein specific cleavage by chymosin. CMP lacks aromatic amino acids, a characteristic that makes it usable as a nutritional supplement for people with phenylketonuria. CMP consists of two nonglycosylated isoforms (aCMP A and aCMP B) and its different glycosylated forms (gCMP A and gCMP B). The most predominant carbohydrate of gCMP is N-acetylneuraminic acid (sialic acid). Here, we developed a CMP purification process based on the affinity of sialic acid for wheat germ agglutinin (WGA). After formation of chitosan beads and adsorption of WGA, the agglutinin was covalently attached with glutaraldehyde. Two matrices with different WGA density were assayed for CMP adsorption. Maximum adsorption capacities were calculated according to the Langmuir model from adsorption isotherms developed at pH 7.0, being 137.0 mg/g for the matrix with the best performance. In CMP reduction from whey, maximum removal percentage was 79% (specifically 33.7% of gCMP A and B, 75.8% of aCMP A, and 93.9% of aCMP B). The CMP was recovered as an aggregate with an overall yield of 64%. Therefore, the matrices developed are promising for CMP purification from cheese whey. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:171-180, 2017.
authors.affiliation: Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956 (1113), Buenos Aires, Argentina.|Instituto de Nanobiotecnología (NANOBIOTEC), UBA, CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín, 956 (1113), Buenos Aires, Argentina.
authors.forename: María F
authors.initials: MF
authors.lastname: Baieli
authors.affiliation: Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956 (1113), Buenos Aires, Argentina.|Instituto de Nanobiotecnología (NANOBIOTEC), UBA, CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín, 956 (1113), Buenos Aires, Argentina.
authors.forename: Nicolás
authors.initials: N
authors.lastname: Urtasun
authors.affiliation: Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Industrias, Ciudad Universitaria (1428), Buenos Aires, Argentina.
authors.forename: María J
authors.initials: MJ
authors.lastname: Martinez
authors.affiliation: Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956 (1113), Buenos Aires, Argentina.|Instituto de Nanobiotecnología (NANOBIOTEC), UBA, CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín, 956 (1113), Buenos Aires, Argentina.
authors.forename: Daniela B
authors.initials: DB
authors.lastname: Hirsch
authors.affiliation: Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Industrias, Ciudad Universitaria (1428), Buenos Aires, Argentina.
authors.forename: Ana M R
authors.initials: AM
authors.lastname: Pilosof
authors.affiliation: Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956 (1113), Buenos Aires, Argentina.|Instituto de Nanobiotecnología (NANOBIOTEC), UBA, CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín, 956 (1113), Buenos Aires, Argentina.
authors.forename: María V
authors.initials: MV
authors.lastname: Miranda
authors.affiliation: Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956 (1113), Buenos Aires, Argentina.|Instituto de Nanobiotecnología (NANOBIOTEC), UBA, CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín, 956 (1113), Buenos Aires, Argentina.
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Cascone
authors.affiliation: Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956 (1113), Buenos Aires, Argentina.|Instituto de Nanobiotecnología (NANOBIOTEC), UBA, CONICET, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín, 956 (1113), Buenos Aires, Argentina.
authors.forename: Federico J
authors.initials: FJ
authors.lastname: Wolman
mesh_terms: D000327:Adsorption; D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002364:Caseins / Q000737:chemistry / Q000302:isolation & purification*; D002417:Cattle; D048271:Chitosan / Q000737:chemistry; D002846:Chromatography, Affinity; D006031:Glycosylation; D008892:Milk / Q000737:chemistry; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D000067796:Whey / Q000737:chemistry; D000067816:Whey Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D000596:Amino Acids; D002364:Caseins; D010446:Peptide Fragments; D000067816:Whey Proteins; C065989:caseinomacropeptide; D048271:Chitosan; D019158:N-Acetylneuraminic Acid
keywords: bovine whey; casein glycomacropeptide; chitosan; chromatography matrix; wheat germ agglutinin