Process for obtaining DNA, RNA, peptides, polypeptides, or protein, by recombinant DNA technique

• US 5,763,192 A
• Filed: 06/05/1995
• Issued: 06/09/1998
• Est. Priority Date: 11/20/1986
• Status: Expired due to Term

• Current Assignee: Stuart Kauffman
• Original Assignee: Ixsys, Inc. (Integral Partners LLC)
• Inventors: Kauffman, Stuart Alan, Ballivet, Marc
• Primary Examiner(s): Achutamurthy, Ponnathapura
• Assistant Examiner(s): PARK, HANKYEL
• Application Number:
  US08/462,637
• Time in Patent Office: 1,100 Days
• Field of Search: 435/7.1, 435/69.1, 435/91.1, 536/23.1
• US Class Current: 435/7.1
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups

Abstract

The present invention is directed to a process for the production of a peptide, polypeptide, or protein having a predetermined property. In accordance with one embodiment, the process begins by producing by way of synthetic polynucleotide coupling, stochastically generated polynucleotide sequences. A library of expression vectors containing such stochastically generated polynucleotide sequences is formed. Next, host cells containing the vectors are cultured so as to produce peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide sequences. Screening or selection is carried out on such host cells to identify a peptide, polypeptide, or protein produced by the host cells which has the predetermined property. The stochastically generated polynucleotide sequence which encodes the identified peptide, polypeptide, or protein is then isolated and used to produce the peptide, polypeptide, or protein having the predetermined property.

Claims

1. A process for the production of a peptide, polypeptide, or protein having a predetermined property, comprising the steps of:

producing by enzymatic or chemical coupling, stochastically generated polynucleotide sequences;

forming a library of expression vectors containing such stochastically generated polynucleotide sequences;

culturing host cells containing the vectors to produce peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide sequences;

carrying out screening or selection on such host cells, to identify a peptide, polypeptide, or protein produced by the host cells having the predetermined property;

isolating a stochastically generated polynucleotide sequence which encodes the identified peptide, polypeptide, or protein;

using the isolated sequence to produce the peptide, polypeptide, or protein having the predetermined property.

2. The process of claim 1, wherein said peptide, polypeptide or protein having said binding property comprises at least one epitope similar to an amino acid sequence of one of the epitopes of a given antigen.

3. The process of claim 1, wherein said peptide, polypeptide or protein having said binding property comprises stimulating or modifying the effects of a biologically active molecule, and screening and/or selection of the clones of transformed host cells producing at least one peptide or polypeptide having this property is carried out by preparing antibodies against that molecule, and utilizing these antibodies so obtained to identify those clones containing those peptides or polypeptides, then by growing the clones thus identified and separating and purifying the peptide or polypeptide produced by these clones, and finally by submitting these peptide(s) or polypeptide(s) to an assay in vitro to verify that it has in fact the capacity to simulate or modify the effects of the said molecule.

4. The process of claims 2 or 3 for the preparation of a vaccine, said process comprising antibodies against a pathogenic agent that are obtained and used to identify those clones producing at least one protein having at least one epitope similar to an amino acid sequence of one of the epitopes of the pathogenic agent, that the corresponding clones of transformed host cells are grown in such a manner as to produce this protein, that the protein is isolated and purified from the cultures of clones of cells and that this protein is used for the production of a vaccine against the pathogenic agent.

5. The process of claim 4 for the preparation of an anti-hepatitis B virus vaccine, said process comprising that at least one capsid protein of the hepatitis B virus is extracted and purified, that this protein is injected into the body of an animal capable of forming antibodies against this protein, that these antibodies are recovered and purified, that these antibodies are used to identify those clones producing at least one protein having at least one epitope similar to one of the epitopes of the hepatitis B virus, that the clones of transformed host cells corresponding to these clones are grown in a manner to produce this protein, that this protein is isolated and purified from these cultures of host cells, and that this protein is used for the production of an anti-hepatitis B virus vaccine.