Matrix screening method
OwnerIan Tomlinson, Lucy J. Holtinventors
Life of this patent
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Abstract
The invention concerns a method which can be used to screen two or more repertoires of molecules against one another and/or to create combinatorial repertoires by combining two or more repertoires. In particular, the invention relates to a method whereby two repertoires of molecules can be screened such that all members of the first repertoire are tested against all members of the second repertoire for functional interactions. Furthermore, the invention relates to the creation and screening of antibody repertoires by combining a repertoire of heavy chains with a repertoire of light chains such that antibodies formed by the all combinations of heavy and light chains can be screened against one or more target ligands.
Claims55
A method for screening a first repertoire of molecules against a second repertoire of molecules to identify those members of the first repertoire which interact with members of the second repertoire, comprising:
- (b) arranging the first and second repertoires to form at least one array, such that all members of the first repertoire are juxtaposed to all members of the second repertoire;
- and
- (b) detecting an interaction between the members of the first and second repertoires, thereby identifying those members of the first repertoire that interact with members of the second repertoire.
A method for screening first, second and third repertoires of molecules against each other to identify those members of the first, second and third repertoires which interact, comprising:
- (a) arranging the first, second and third repertoires to form at least one array, such that all members of the first, second and third repertoires are juxtaposed;
- and
- (b) detecting an interaction between the members of the first, second and third repertoires, thereby identifying those members of the first, second and third repertoires that interact.
A method for creating a combinatorial library of two-chain polypeptides, each member of which library comprises one member of a first repertoire of single chain polypeptides and one member of a second repertoire of single chain polypeptides, which method comprises the step of arranging the first and second repertoires of single chain polypeptides, such that all members of the first repertoire intersect all members of the second repertoire, thereby generating at their intersections all combinations of functional two-chain polypeptides, thereby creating a combinatorial library of two-chain polypeptides.
A method for creating a combinatorial library of three-chain polypeptides, each member of which library comprises one member of a first repertoire of single chain polypeptides, one member of a second repertoire of single chain polypeptides, and one member of a third repertoire of single chain polypeptides, which method comprises:
- (a) arranging the first, second and third repertoires of single chain polypeptides, such that all members of each repertoire intersect all members of the other two repertoires, thereby generating at their intersections all combinations of functional three-chain polypeptides, thereby creating a combinatorial library of three-chain polypeptides.
A method for screening a first repertoire of molecules against a second repertoire of molecules to identify those members of the first repertoire which do not interact with members of the second repertoire, the method comprising:
- (a) arranging the first and second repertoires, such that all members of the first repertoire are juxtaposed with all members of the second repertoire;
- and
- (b) identifying those members of the first and second repertoires that do not interact with one another.
A method for screening a first repertoire of molecules against a second repertoire of molecules to identify members of the first and second repertoires whose interactions with one another are dependent on the presence or absence of a third molecule or set of molecules, comprising:
- (a) arranging the first and second repertoires, such that all members of the first repertoire are juxtaposed with all members of the second repertoire;
- and
- (b) detecting interactions between members of the first repertoire and members of the second repertoire in the presence of different concentrations of the third molecule or set of molecules, such that members of the first and second repertoires whose interactions with one another are dependent on the presence or absence of the third molecule or set of molecules are identified.
An apparatus for drawing, on a medium, lines of matter comprising members of a first, second and, optionally third repertoire of molecules.
An apparatus which comprises intersecting channels or tubes along which members of a first, second and, optionally third repertoire of molecules can pass.
A method for optimizing conditions for a biological interaction, which method comprises creating all combinations of two or more different sets of variable parameters at the intersections of two or more different sets of intersecting lines, channels or tubes, and assaying the biological interaction, thereby optimizing the conditions for the biological interaction.
A method for screening a first and a second repertoire of enzymes to identify those members of the first repertoire and those members of the second repertoire which together participate in a two or more step enzymatic reaction that creates a given product from a given substrate, which method comprises:
- (a) arranging the first and second repertoires to form at least one array, such that all members of the first repertoire are juxtaposed to all members of the second repertoire;
- and
- (b) detecting the formation of the given product at the intersections of the members of the first and second repertoires, thereby identifying those members of the first and second repertoires which together participate in a two or more step enzymatic reaction that creates the given product from the given substrate.
A method for screening a plurality of cellular populations against a plurality of viral populations to identify those viral populations among the plurality of viral populations that infect cellular populations among the plurality of cellular populations, which method comprises:
- (a) arranging the plurality of cellular populations and the plurality of viral populations to form at least one array, such that all the different cellular populations are juxtaposed with all the viral populations;
- and
- (b) detecting viral infection in the plurality of cellular populations, thereby identifying those viral populations among the plurality of viral populations that infect cellular populations among the plurality of cellular populations.
A method for screening a plurality of different cellular fractions against one another to identify those cellular fractions that contain components which interact with components in the other cellular fractions, which method comprises:
- (a) arranging the plurality of cellular fractions to form at least one array, such that all the different cellular fractions are juxtaposed to one another;
- and
- (b) detecting the interaction of different cellular fractions at sites where the different cellular fractions are juxtaposed, thereby identifying those cellular fractions that contain components which interact with components in the other cellular fractions tested.
A method for screening a plurality of different cellular populations against one another to identify those cellular populations that interact with the other cellular populations, which method comprises:
- (a) arranging the plurality of cellular different populations to form at least one array, such that all the different cellular populations are juxtaposed to one another;
- and
- (b) detecting the interaction of different cellular fractions at sites where the different cellular fractions are juxtaposed, thereby identifying those cellular populations that interact with the other cellular populations tested.
A method for screening a peptide repertoire against the same peptide repertoire to identify those members of the peptide repertoire that interact with other members of the peptide repertoire, which method comprises:
- (a) arranging the members of the peptide repertoire to form at least one array, such that all the members of the peptide repertoire are juxtaposed to one another;
- and
- (b) detecting the interaction of different members of the peptide repertoire at sites where the different members are juxtaposed, whereby those members of the peptide repertoire that interact with other members of the same peptide repertoire are identified.
A method for screening a polypeptide repertoire against the same polypeptide repertoire, in order to identify those members of the polypeptide repertoire that interact with other members of the polypeptide repertoire, which method comprises:
- (a) arranging the members of the polypeptide repertoire to form at least one array, such that all the members of the polypeptide repertoire are juxtaposed to one another;
- and
- (b) detecting the interaction of different members of the polypeptide repertoire at sites where the different members are juxtaposed, thereby identifying those members of the polypeptide repertoire that interact with other members of the polypeptide repertoire.
A method for creating a combinatorial library consisting of all members of a first repertoire of polypeptides paired with all members of a second repertoire of polypeptides, which method comprises:
- (a) arranging a repertoire of host cells containing a plurality of nucleotide sequences encoding a first repertoire of polypeptide members, and a plurality of nucleotide sequences encoding a second repertoire of polypeptide members to create an array, such that cells containing nucleotide sequences encoding all members of the first repertoire intersect with nucleotide sequences corresponding to all members of the second repertoire;
- and
- (b) transforming the cells containing the nucleotide members of the first repertoire with the nucleotide sequences that encode the members of the second repertoire where the two repertoires intersect;
- and
- (c) expressing the nucleotide sequences to produce the corresponding polypeptides of the first and second repertoires;
- thereby creating a combinatorial library consisting of all members of the first repertoire of polypeptides paired with all members of the second repertoire of polypeptides.
A method for creating a combinatorial library consisting of all members of a first repertoire of polypeptides paired with all members of a second repertoire of polypeptides, which method comprises:
- (a) arranging a repertoire of host cells containing a plurality of nucleotide sequences encoding a first repertoire of polypeptide members, and a plurality of viruses containing a plurality of nucleotide sequences encoding a second repertoire of polypeptide members to create an array, such that cells containing all nucleotide members of the first repertoire intersect with viruses containing all nucleotide members of the second repertoire;
- (b) infecting the cells containing the nucleotide members of the first repertoire with the viruses that contain the nucleotide members of the second repertoire where the two repertoires intersect;
- and
- (c) expressing the nucleotide sequences to produce the corresponding polypeptides of the first and second repertoires, thereby creating a combinatorial library consisting of all members of the first repertoire of polypeptides paired with all members of the second repertoire of polypeptides.
A method for creating a yeast two hybrid library consisting of all members of a first repertoire of polypeptides paired with all members of a second repertoire of polypeptides, which method comprises:
- (a) arranging yeast cells containing a plurality of nucleotide sequences encoding a first repertoire of polypeptide members, and yeast cells containing a plurality of nucleotide sequences encoding a second repertoire of polypeptide members to create an array, such that yeast cells containing all nucleotide members of the first repertoire intersect with yeast cells containing all nucleotide members of the second repertoire;
- (b) allowing the yeast cells containing the members of the first repertoire to mate with the yeast cells containing the members of the second repertoire where the two repertoires intersect;
- and
- (c) expressing the nucleotide sequences to produce the corresponding polypeptides of the first and second repertoires, thereby creating a yeast two hybrid library consisting of all members of a first repertoire of polypeptides paired with all members of a second repertoire of polypeptides.
Dependencies are inferred from the claim text.