Isolation and production of catalytic antibodies using phage technology
First Claim
Patent Images
1. A method for producing catalytic antibodies displayed on phage comprising the steps of:
- (a) generating a gene library of antibody-derived domains;
(b) inserting coding for said domains into a phage expression vector;
(c) isolating said catalytic antibodies; and
(d) wherein said phage expression vector incorporates therein a histidine peptide in tandem with a myc peptide.
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Abstract
Disclosed and claimed are methods for producing catalytic antibodies, including human catalytic antibodies, from bacteriophage. The methods require the cloning, selection, screening, and isolation of catalytic antibodies. Also disclosed and claimed are the products themselves, the catalytic antibodies, made from the phage technology. In addition, catalytic antibodies produced from the phage technology and useful in prodrug activation are disclosed and claimed. And finally, the invention also understands the production of catalytic antibodies to phosphonates.
79 Citations
26 Claims
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1. A method for producing catalytic antibodies displayed on phage comprising the steps of:
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(a) generating a gene library of antibody-derived domains; (b) inserting coding for said domains into a phage expression vector; (c) isolating said catalytic antibodies; and (d) wherein said phage expression vector incorporates therein a histidine peptide in tandem with a myc peptide. - View Dependent Claims (2, 3, 4, 5, 23)
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6. A method for isolating catalytic antibodies displayed on phage comprising the steps:
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(a) preparing an antigen; (b) immunizing with said antigen; (c) generating a library of VH and VL domains from said immunized animal; (d) cloning said VH and VL domains into a phage expression vector to generate phage display antibodies; (e) selecting phage display antibodies which bind specifically to said antigen; (f) screening said selected phage display antibodies for catalytic activity to substrate; (g) isolating said catalytic antibodies; and (h) wherein said phage expression vector incorporates therein a histidine peptide in tandem with a myc peptide. - View Dependent Claims (7, 8, 9, 10, 11)
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12. A method for isolating catalytic antibodies displayed on phage comprising the following steps:
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(a) preparing an antigen; (b) generating a library of VH and VL domains; (c) cloning said VH and VL domains into a phage expression vector to generate phage display antibodies; (d) selecting phage display antibodies which bind specifically to said antigen; (e) screening said selected phage display antibodies for catalytic activity to substrate; (f) isolating said catalytic antibodies; and (g) wherein said phage expression vector incorporates therein a histidine peptide in tandem with a myc peptide. - View Dependent Claims (13, 14, 15, 16, 17)
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18. A method for producing catalytic antibodies displayed on phage through chain shuffling comprising the following steps:
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(a) combining a library of VL genes with VH genes to form a chain shuffled library; (b) cloning the shuffled chain; (c) expressing said chain shuffled antibody on phage; (d) selecting against an antigen; (e) screening for catalytic activity; and (f) wherein said phage incorporates therein a histidine peptide in tandem with a myc peptide. - View Dependent Claims (24)
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19. A method for producing catalytic antibodies displayed on phage through CDR shuffling comprising the following steps:
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(a) isolating VL and VH genes; (b) isolating a library of CDR regions; (c) recombining said VL and VH genes with said library of CDR regions to produce a CDR shuffled library; (d) cloning the CDR shuffled library; (e) expressing said CDR shuffled library on phage; (f) selecting against an antigen; (g) screening for catalytic activity; and (h) wherein said phage incorporates therein a histidine peptide in tandem with a myc peptide. - View Dependent Claims (25)
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20. A method for producing catalytic antibodies displayed on phage through imprinting comprising the following steps:
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(a) selecting a set of antibodies; (b) isolating a set of VH and a set of VL genes from said antibodies; (c) combining said set of VH with a library of VL and combining said set of VL with a library of VH to form two combinatorial libraries; (d) cloning said combination libraries; (e) expressing said libraries on phage; (f) selecting against an antigen; (g) isolating selected libraries of VH and VL genes; (h) combining said libraries of VH and VL genes; (i) cloning said combined libraries; (j) expressing said combined libraries on phage; (k) reselecting against an antigen; (l) screening for catalytic activity; and (m) wherein said phage incorporates therein a histidine peptide in tandem with a myc peptide. - View Dependent Claims (26)
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21. A method for enhancing the rate of cleavage or formation of a specific bond within a molecule in vivo which comprises introducing into an animal an effective amount of a phage-derived catalytic antibody;
- and wherein said phage incorporates therein a histidine peptide in tandem with a myc peptide.
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22. A method for in vivo activation of a prodrug comprising:
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(a) introducing a prodrug into a patient, said prodrug having a chemical bond therein which upon cleavage releases the active form of said drug; (b) introducing into said patient an effective amount of a phage-derived catalytic antibody capable of cleaving said bond in said prodrug; and (c) wherein said phage incorporates therein a histidine peptide in tandem with a myc peptide.
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Specification