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US 6,207,390 B1Granted
Methods for the use of reduced affinity streptavidin
AssigneeTrustees of Boston University
InventorsCharles R. Cantor, Takeshi Sano
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods for contacting biological targets using a mutated streptavidin protein having a reduced affinity for biotin.
Claims16
A method for contacting a target, comprising a) providing a target to be biotinylated and biotinylation means;
- b) biotinylating said target with said biotinylation means under conditions such that a heterogeneous mixture is created, said heterogeneous mixture comprising target and biotinylated target;
- and c) contacting said heterogeneous mixture with a solid support, said solid support comprising a reduced-affinity streptavidin protein, wherein said reduced-affinity streptavidin protein has an amino acid sequence that is different from the amino acid sequence of wild-type streptavidin (SEQ ID NO:5), and wherein the difference between said reduced-affinity streptavidin protein and said wild-type streptavidin (SEQ ID NO:5) comprises deletions of residues 113-120 from the 16-133 fragment of said wild-type streptavidin.
A method for contacting a target, comprising a) providing a target to be biotinylated and biotinylation means;
- b) biotinylating said target with said biotinylation means under conditions such that a heterogeneous mixture is created, said heterogeneous mixture comprising target and biotinylated target;
- and c) contacting said heterogeneous mixture with a solid support, said solid support comprising a reduced-affinity streptavidin protein, wherein the difference between said reduced-affinity streptavidin protein and wild-type streptavidin (SEQ ID NO:5) comprises a phenylalanine substituted for a tryptophan at positions 79, 92, 108, or 120 of said wild-type streptavidin (SEQ ID NO:5) such that said reduced-affinity streptavidin protein displays reduced-affinity for biotin.
A method for contacting a target, comprising a) providing a target to be biotinylated and biotinylation means;
- b) biotinylating said target with said biotinylation means under conditions such that a heterogeneous mixture is created, said heterogeneous mixture comprising target and biotinylated target;
- and c) contacting said heterogeneous mixture with a solid support, said solid support comprising a reduced-affinity streptavidin protein, wherein said reduced-affinity streptavidin protein has an amino acid sequence that is different from the amino acid sequence of SEQ ID NO:1, and wherein the difference between said reduced-affinity streptavidin protein and said SEQ ID NO:1, comprises deletions of residues 113-120 from the 16-133 fragment of said amino acid sequence of SEQ ID NO:
- 1.
A method for contacting a target, comprising a) providing a target to be biotinylated and biotinylation means;
- b) biotinylating said target with said biotinylation means under conditions such that a heterogeneous mixture is created, said heterogeneous mixture comprising target and biotinylated target;
- and c) contacting said heterogeneous mixture with a solid support, said solid support comprising a reduced-affinity streptavidin protein, wherein the difference between said reduced-affinity streptavidin protein and the amino acid sequence of SEQ ID NO:1 comprises a phenylalanine substituted for a tryptophan at positions 79, 92, 108, or 120 of said amino acid sequence of SEQ ID NO:
- 1 such that said reduced-affinity streptavidin protein displays reduced-affinity for biotin.
Dependencies are inferred from the claim text.