Purification And Identification Of The HER2Protein In Breast Cancer Tissue

Background：Malignant tumors have become the first threat to human lifeand health. In order to control the development of tumor better, explore thetherapeutic methods of cancer and monitor the tumor development and prognosis,people have made a lot of research work on tumor, and established a series ofreference indicators to detect tumors. In the study of a wide variety of tumor hasdiscovered a proto-oncogene HER2. HER2further study found that the highproto-oncongene expression of HER2protein product, also showed a highexpression in breast cancer, cervical cancer, ovarian cancer, and cancerous tissue.HER2protein, as a major tumor surface marker protein, especially in breast cancer,has been used as an important indicator in clinical. Therefore, the blood and tissuesimmunochemical detection of HER2protein has also been developing rapidly, andthus increasing demand for its standard antigen, so that to establish a stable anddfficient preparation process highlights the importance of HER2protein.Objective： Establish a complete set of method that can extract the HER2protein from breast tissue, and prepare high purity HER2protein. The high purityHER2protein will create conditions for preparation of anti-HER2antibody and thedevelopment of appropriate immunological detection methods and related vaccines.Methods: Used sonication prepared breast cancer tissue homogenates,centrifuged, ammonium sulfate precipitation and ultrafiltration method crude extractHER2protein. Then, with the methods of molecular screening chromatography, ionexchange chromatography, purified the HER2protein from the crude extractedprotein. Identified the purified protein by SDS-PAGE, Western-blot and ELISA.Results: Established the method of extracted and purified HER2protein fromhuman breast cancer tissues. The molecular size of the purified protein is about185kD, the HER2protein which have immune activity and it is more than95%purity. Conclusion: Through this research, establish the purification technology, highpurity, with immune activity HER2protein was prepared from human breast cancertissue.