Method of preparing antibodies to characterize oncogenes

A method of producing antibody capable of detecting carcinogenesis caused by mutation of a proto-oncogene into an oncogene, comprising:

• a. isolating a segment of DNA, the nucleotide sequence of which is different in said proto-oncogene from the nucleotide sequence of said oncogene, the difference comprising a nucleotide substitution;
• b. determining the nucleotide sequence of the proto-oncogene and the nucleotide sequence of the oncogene;
• c. deducing the amino acid sequence of the protein encoded by the proto-oncogene and the amino acid sequence of the protein encoded by the oncogene;
• d. determining the segment of the amino acid sequence which is different in the two encoded proteins and is critical for their difference in function;
• e. synthesizing an oligopeptide which includes an amino acid sequence present in the segment of the protein encoded by the oncogene but not present in the corresponding segment of the protein encoded by the proto-oncogene and critical for the difference in function of the two proteins;
• f. synthesizing an oligopeptide of the corresponding region of the oncogene-encoded protein;
• g. immunizing a host with the oligopeptide corresponding to an amino acid sequence encoded by said oncogene, under conditions whereby cells of said host produce antibody against said segment.

A method of producing antibodies specific for a protein encoded by an oncogene or a protein encoded by its corresponding proto-oncogene, but not for both, comprising the steps of:

• a. isolating a nucleotide sequence present in the oncogene or present in its corresponding proto-oncogene, but not in both the oncogene and the corresponding proto-oncogene;
• b. producing an antigen encoded by the nucleotide sequence present in the oncogene or by the nucleotide sequence present in the corresponding proto-oncogene;
• and
• c. immunizing an appropriate host with the antigen encoded by the nucleotide sequence present in the oncogene or with the antigen encoded by the nucleotide sequence present in the corresponding proto-oncogene, under conditions whereby the host produces antibodies specific for the antigen.

A method of producing antibodies capable of determining whether a proto-oncogene has been mutated to its corresponding oncogene, comprising the steps of:

• a. producing a DNA fragment having the nucleotide sequence of DNA of the proto-oncogene which is mutated to produce the nucleotide sequence of the oncogene;
• b. producing a protein encoded by the nucleotide sequence of the oncogene, the protein capable of causing, upon introduction into an appropriate host, production of antibodies specific for protein encoded by the oncogene, but not for both;
• and
• c. immunizing a host, under conditions appropriate for producing antibodies.