Affinity precipitation of proteins.
Affinity precipitation is being studied as a technique to be introduced at an early stage of downstream processing for the selective isolation of proteins. The technique utilizes a heterobifunctional ligand, which, in addition to having affinity for the target protein(s), possesses another function for controlling precipitation. The latter component is comprised of a polymer which can be made reversibly soluble and insoluble by altering a specific parameter such as pH or temperature. Different polymers of natural and synthetic origin have been used for this purpose. The soluble form of the ligand is used for the affinity binding step and precipitation is induced for obtaining separation of the affinity complex. Some of the polymers used in this laboratory include chitosan, alginate, Eudragit S-100 (copolymer of methacrylic acid and methyl methacrylate) and polyethyleneimine. Chitosan and alginate served as natural ligands for wheat germ agglutinin and pectinase, respectively. The aromatic dye Cibacron Blue 3GA coupled to Eudragit S 100 and polyethyleneimine way used for the affinity precipitation of some model enzymes such as lactate dehydrogenase and alcohol dehydrogenase. As prior removal of cell debris, etc., is essential for affinity precipitation, the possibility of integration of the technique with extraction in aqueous two-phase systems was also demonstrated.
Gupta
M N
MN
Department of Biotechnology, Lund University, Sweden.
Kaul
R
R
Guoqiang
D
D
Dissing
U
U
Mattiasson
B
B
D011232:Chemical Precipitation; D008024:Ligands; D011108:Polymers; D011109:Polymethacrylic Acids; D011506:Proteins / Q000302:isolation & purification*
D008024:Ligands; D011108:Polymers; D011109:Polymethacrylic Acids; D011506:Proteins; C038300:methylmethacrylate-methacrylic acid copolymer