Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1992-04-15
1994-05-31
Hill, Jr., Robert J.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 737, 435 78, 435 732, 935 79, G01N 3353, C12Q 102, C12N 1510
Patent
active
053169220
ABSTRACT:
This invention provides for improved means to produce binding specific proteins using concatamers of semi-randomly generated oligonucleotides inserted into genes encoding the external domains of bacterial outer membrane proteins. The genes are induced to express and the bacteria are then screened for the ability to bind to predetermined compositions. Those clones carrying the desired binding protein are isolated, cultured and the protein purified. Increased avidity of the binding specific proteins are achieved by reisolating the oligonucleotides which conferred binding affinity and mixing them with new semi-randomly generated oligonucleotides to generate a population enriched for oligonucleotides that had previously conferred to bacteria the desired binding affinity. The enriched population of oligonucleotides is then religated to the gene encoding the external domain of the bacterial protein, the gene is inserted into a bacterial cell and the cells analyzed for increased avidity for the predetermined composition.
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Devlin, J. J., et al., "Random Peptide Libraries: A Source of Specific Protein Binding Molecules", Science 249:404-406 (1990).
Clackson, T., et al., "Making Antibody Fragments Using Phage Display Libraries", Nature 352:624-628 (1991).
Cwirla, S. E., et al., "Peptides on Phage: A Vast Library of Peptides for Identifying Ligands", Proc. Natl. Acad. Sci. USA 87:6378-6382 (1990).
Scott, J. K. and Smith, G. P., "Searching for Peptide Ligands with an Epitope Library", Science 249:386-390 (1990).
Brown Stanley
Court Donald
Hill Jr. Robert J.
Spector Lorraine M.
The United States of America as represented by Department of Hea
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