RNA-antibody fusions and their selection
First Claim
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1. A molecule comprising a ribonucleic acid covalently bonded through an amide bond to an antibody, wherein said antibody is encoded by said ribonucleic acid.
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Abstract
Described herein are methods and reagents for the selection of protein molecules that make use of RNA-protein fusions.
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Citations
12 Claims
- 1. A molecule comprising a ribonucleic acid covalently bonded through an amide bond to an antibody, wherein said antibody is encoded by said ribonucleic acid.
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2. A molecule comprising a ribonucleic acid covalently bonded to an antibody, said antibody being entirely encoded by said ribonucleic acid.
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9. An RNA-antibody fusion selected by the method comprising the steps of:
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a) providing a population of candidate RNA molecules, each of which comprises a translation initiation sequence and a start codon operably linked to a candidate antibody coding sequence and each of which is operably linked to a peptide acceptor at the 3′
end of said candidate antibody coding sequence;
b) in vitro or in situ translating said candidate antibody coding sequences to produce a population of candidate RNA-antibody fusions;
c) contacting said population of candidate RNA-antibody fusions with a target antigen; and
d) selecting an RNA-antibody fusion that binds to said target antigen.
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10. An RNA-antibody fusion selected by the method comprising the steps of:
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a) providing a population of candidate RNA molecules, each of which comprises a translation initiation sequence and a start codon operably linked to a candidate antibody coding sequence and each of which is operably linked to a peptide acceptor at the 3′
end of said candidate antibody coding sequence;
b) in vitro translating said candidate antibody coding sequences to produce a population of candidate RNA-antibody fusions;
c) contacting said population of candidate RNA-antibody fusions with a target antigen; and
d) selecting an RNA-antibody fusion that binds to said target antigen.
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11. An RNA-antibody fusion comprising an antibody portion having an altered binding affinity or specificity for a target antigen relative to the binding affinity or specificity of a reference antibody, said RNA-antibody fusion selected by the method comprising the steps of:
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a) producing a population of candidate RNA molecules, each having a candidate antibody coding sequence which differs from said reference antibody coding sequence, said RNA molecules each comprising a translation initiation sequence and a start codon operably linked to said candidate antibody coding sequence and each being operably linked to a peptide acceptor at the 3′
end;
b) in vitro or in situ translating said candidate antibody coding sequences to produce a population of candidate RNA-antibody fusions;
c) contacting said population of candidate RNA-antibody fusions with said target antigen; and
d) selecting an RNA-antibody fusion, the antibody portion of which has a binding affinity or specificity for said target antigen that is altered relative to the binding affinity or specificity of said reference antibody for said target antigen.
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12. An RNA-antibody fusion comprising an antibody portion having an altered binding affinity or specificity for a target antigen relative to the binding affinity or specificity of a reference antibody, said RNA-antibody fusion selected by the method comprising the steps of:
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a) producing a population of candidate RNA molecules, each having a candidate antibody coding sequence which differs from said reference antibody coding sequence, said RNA molecules each comprising a translation initiation sequence and a start codon operably linked to said candidate antibody coding sequence and each being operably linked to a peptide acceptor at the 3′
end;
b) in vitro translating said candidate antibody coding sequences to produce a population of candidate RNA-antibody fusions;
c) contacting said population of candidate RNA-antibody fusions with said target antigen; and
d) selecting an RNA-antibody fusion, the antibody portion of which has a binding affinity or specificity for said target antigen that is altered relative to the binding affinity or specificity of said reference antibody for said target antigen.
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Specification