Chemistry: molecular biology and microbiology – Carrier-bound or immobilized enzyme or microbial cell;... – Enzyme or microbial cell is immobilized on or in an organic...
Patent
1996-07-24
2000-04-11
Naff, David M.
Chemistry: molecular biology and microbiology
Carrier-bound or immobilized enzyme or microbial cell;...
Enzyme or microbial cell is immobilized on or in an organic...
435 681, 435 691, 435 697, 435 701, 435 711, 435395, 435803, 435815, 435 712, 4351723, 435178, 4353201, 530413, 530813, 530814, 436529, 436530, C12N 1112, C12N 1500, C12P 2106, A23J 100
Patent
active
060487151
ABSTRACT:
A two-phase partition system is provided for affinity separation of a composition containing a polysaccharide binding peptide from a mixture such as a fermentation broth. The peptide may be from an enzyme and lacking in polysaccharidase activity such as the binding domain of cellulase that binds to cellulose. The system contains a phase-forming oligosaccharide polymer such as a cellulose derivative to which the peptide binds with a Ka of 10.sup.3 M to 10.sup.7 M, and a phase inducing agent such as a polyethylene glycol polymer, or a salt present at sufficiently high concentration to induce phase separation. If the oligosaccharide polymer is thermoseparating, phase separation can be induced by heating. Using the system involves contacting a composition containing the peptide such as a fusion protein with the system, partitioning the composition into a phase containing the oligosaccharide polymer by binding to the polymer and recovering the polymer containing the bound composition. The peptide or a fusion protein containing the peptide can be contacted with a cell having a carbohydrate residue to which the peptide binds to form a complex, and the complex is separated with the system to produce a bound cell composition. The peptide may be linked through a protease recognition sequence to a macromolecule such as an enzyme, a hormone or an antibody, and the macromolecule can be removed by using a protease to cleave the recognition sequence.
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Haynes Charles A.
Kilburn Douglas G.
Tomme Peter
Naff David M.
Univ. of British Columbia
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