Materials and methods for detection and treatment of breast cancer
a breast cancer and material technology, applied in the field of breast cancer detection and/or treatment, can solve the problems of high risk of breast cancer, limited brca1 as a cancer marker, and breast cancer is a leading cause of death in women, so as to reduce the expression of the target protein, and reduce the risk of breast cancer. the effect of in vivo expression
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example 1
Identification of Breast Cancer Markers
[0115] To identify markers for breast cancer, the sera of individuals with breast cancer were compared to the sera of normal individuals by surface-enhanced laser desorption and ionization (SELDI) mass spectrometry. Briefly, 0.5 mL aliquots of sera harvested from the individuals were thawed. Then, 1 μL of a 1 mg / mL solution of soybean trypsin inhibitor (SBTI) and 1 μL of a 1 mg / mL solution of leupeptin were added to each aliquot. To remove lipids, 350 μL of 1,1,2-trifluorotrichloroethane was added to each sample. The samples then were vortexed for five minutes and centrifuged in a microcentrifuge for five minutes at 4° C. The resulting supernatants were applied a 1 mL column of agarose coupled to protein G (Hitrap Protein G column, Pharmacia and Upjohn, Peapack, N.J.) to remove immunoglobulin proteins. The column then was rinsed with 3 mL of 50 mM sodium phosphate, pH 7.0, with SBTI and leupeptin (“binding buffer”), and the resulting flowthrou...
example 2
Sequencing of Breast Cancer Marker Proteins
[0124] Breast cancer-associated proteins based upon the biochemical and mass spectrometry data provided above may be better characterized using well-known techniques. For example, samples of the serum can be fractionated using, for example, column chromatography and / or electrophoresis, to produce purified protein samples corresponding to each of the proteins identified in Table 1. The sequences of the isolated proteins can then be determined using conventional peptide sequencing methodologies (see Examples 5 and 6). It is appreciated that the skilled artisan, in view of the foregoing disclosure, would be able to produce an antibody directed against any breast cancer-associated protein identified by the methods described herein. Moreover, the skilled artisan, in view of the foregoing disclosure, would be able to produce nucleic acid sequences that encode the fragments described above, as well as nucleic acid sequences complementary thereto....
example 3
Production of Antibodies Which Bind Specifically to Breast Cancer-Associated Proteins
[0125] Once identified, a breast cancer-associated protein may be detected in a tissue or body fluid sample using numerous binding assays that are well known to those of ordinary skill in the art. For example, as discussed above, a breast cancer-associated protein may be detected in either a tissue or body fluid sample using an antibody, for example, a monoclonal antibody, which binds specifically to an epitope disposed upon the breast cancer-associated protein. In such detection systems, the antibody preferably is labeled with a detectable moiety.
[0126] Provided below is an exemplary protocol for the production of an anti-breast cancer-associated monoclonal antibody. Other protocols also are envisioned. Accordingly, the particular method of producing antibodies to target proteins is not envisioned to be an aspect of the invention.
[0127] Balb / c by J mice (Jackson Laboratory, Bar Harbor, Me.) are ...
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