Method for the selecting of genes encoding catalytic antibodies
OwnerClaude G. Davis, Gary R. Fabianinventors
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Abstract
The present invention describes methods of screening for or selecting a catalytic antibody effective to cleave a target peptide. In particular, in the selection method of the present invention a phage gene is selected that encodes a gene product necessary for the production of a phage. The phage, carrying the modified gene is introduced into a host. Also, a library of rearranged immunoglobulin genes in a cloning vector is introduced into host cells. The host cells are grown under conditions in which the immunoglobulin genes are expressed in the host cells. The presence of antibodies capable of cleaving the target peptide is identified on the basis of production of phage.
Claims8
A method of selecting genes encoding catalytic antibodies which are capable of cleaving a selected target peptide, comprising
- introducing into bacterial cells (i) a library of rearranged immunoglobulin genes in a cloning vector capable of expressing immunoglobulin genes in the cloning vector, under suitable expression conditions, and (ii) a phage vector bearing a phage gene encoding a gene product necessary for the production of infectious phage, where said gene is modified by introducing the target peptide coding sequence into the gene such that the resulting gene product inhibits production of infectious phage, and where cleavage of said target peptide results in an active gene product that allows production of infectious phage,
- growing the bacterial cells under conditions in which the immunoglobulin genes are expressed in the bacterial cells,
- screening the bacterial cells for production of infectious phage, and
- isolating the immunoglobulin genes associated with the infectious phage.
A method of producing a catalytic antibody effective to cleave a target peptide sequence, comprising
- introducing into bacterial cells (i) a library of rearranged immunoglobulin genes in a cloning vector capable of expressing immunoglobulin genes in the cloning vector, under suitable expression conditions, and (ii) a phage vector bearing a phage gene encoding a gene product necessary for the production of infectious phage, where said gene is modified by introducing the target peptide coding sequence into the gene such that the resulting gene product inhibits production of infectious phage, and where cleavage of said target peptide results in an active gene product that allows production of infectious phage,
- growing the bacterial cells under conditions in which the immunoglobulin genes are expressed in the bacterial cells,
- screening the bacterial cells for production of infectious phage,
- isolating the immunoglobulin genes associated with the infectious phage, and
- expressing the isolated immunoglobulin genes in a suitable expression system.
Dependencies are inferred from the claim text.