Promotion of high specificity molecular assembly
AssigneeThe Board of Trustees of the Leland Stanford Jr. University
InventorBrian W. Pontius
Life of this patent
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Abstract
This invention relates to an improvement in promoting the rate of association for high specificity binding pairs used in a variety of industrial, research and medical applications. These pairs include enzyme/substrate, complementary polynucleotide and antibody/antigen combinations. In one specific embodiment, this invention relates to the acceleration of nucleic acid hybridization by heterogeneous nuclear ribonucleoproteins [hnRNPs]. In another specific embodiment, this invention relates to the acceleration of nucleic acid hybridization by a cationic detergent.
Claims20
A composition comprising complementary members of a primary binding pair covalently bound to complementary members of a secondary binding pair where the members of the secondary pair have an association rate constant which exceeds the association rate constant of the primary binding pair members and wherein complementary primary binding pair members are bound to complementary secondary binding pair members such that the binding between secondary binding pair members increases the rate of association for the primary binding pair members.
A method for increasing the association rate constant for members of primary binding pairs in the absence of aggregate formation by the primary binding pairs;
- said method comprising (a) combining the primary binding pair members in an aqueous solution wherein at least one of the primary binding pair members is covalently bound to a multivalent association rate enhancer;
- and, (b) incubating the binding pairs under conditions which permit association of the binding pairs and the multivalent rate enhancers.
A method for increasing the association rate constant for members of a primary binding pair in an enzyme reaction wherein primary bind pair consists of an enzyme and substrate, said method comprising:
- (a) attaching a polymer to the enzyme, said polymer binding with high probability to the substrate such that the polymer and substrate have a k.sub.a larger than the k.sub.a of the enzyme and substrate said k.sub.a being measured when the polymer and enzymes are physically apart;
- and
- (b) placing the members in an aqueous solution under conditions which permit binding between members of the primary binding pair.
A method for increasing the association rate constant for members of primary binding pairs, said method comprising
- (a) covalently attaching complementary members of secondary binding pairs to complementary members of primary pairs where members of the secondary binding pairs have a k.sub.a larger for each other than the k.sub.a of the members of the primary binding pairs for each other and where the members of the secondary binding pairs are attached in a manner permitting them to associate with their respective complementary members thereby increasing the association rate constant for the primary binding pair members;
- and,
- (b) placing the members in an aqueous solution under conditions which permit binding between members of the primary binding pairs;
- wherein said primary binding pairs are substantially apart prior to initiating step (b).
Dependencies are inferred from the claim text.