Polysaccharide binding fusion proteins and conjugates

Details Polysaccharide binding fusion proteins and conjugates Polysaccharide binding fusion proteins and conjugates

1997-01-24

1999-10-05

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 4, 435 691, 435 6952, 435 697, 435 711, 4351723, 435177, 435195, 435200, 435209, 435803, 435178, 436530, 530402, 530808, 530814, 530413, 935 11, 935 14, C12N 1112, C12N 1500, C12P 2100

Patent

active

059622890

ABSTRACT:
Fusion proteins or conjugates are provided containing an amino acid sequence having a substrate binding region of a polysaccharidase such as cellulase that binds to a .beta.-1,4-glycan matrix such as cellulose. The substrate binding region is essentially without polysaccharidase activity. In the fusion protein, the substrate binding region is fused or chemically linked to a polypeptide such as an enzyme, a hormone, an immunoglobulin or a protein dye. By contacting the fusion protein with a .beta.-1,4-glycan matrix, the substrate binding region binds to the matrix to immobilize the polypeptide on the matrix. The polypeptide or fusion protein can be removed from the matrix with a protease acting on a protease recognition sequence or with a solution having a low ionic strength or high pH. In the conjugate, the substrate binding region is joined such as by covalent bonding to a non-protein chemical moiety such as a dye, chromophore, fluorescor, radionuclide or enzyme co-factor. By contacting the conjugate with a .beta.-1,4-glycan matrix, the substrate binding region binds to the matrix to immobilize the chemical moiety on the matrix. The conjugate or chemical moiety can be removed with a protease acting on a protease recognition sequence or with a solution having a low ionic strength or high pH.

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