Gene Expression, Antibody Prepration And Tissues Distribution Investigation Of NY-ESO-1

Objective: Tumor-targeting immunotherapy is a new strategy for treating the malignanttumor. And the key factor for it is identification of tumor-specific antigen. Recent studies have described that the cancer-testis (CT) antigens, which express in limited testis and ovaries primarily, are expressed in a wide variety of cancers. NY-ESO-1, as a member of CT family, has an advantage to induce both humoral and cellular immune responses in NY-ESO-1~+ patients. So, upon to these characterizations, NY-ESO-1 is nearly an ideal candidate target antigen for cancer immunotherapy. We expressed human NY-ESO-1 by recombination with GST in E.coli and purified this fusion protein by GST affinity chromgraphy. The polyclonal as well as monoclonal antibodies were prepared by immunizing Balb/c mouse. Based on our preliminary data and promising prospection, NY-ESO-1 will act as an important tumor-vaccine candidate for cancer therapy.Methods: The coding region of human NY-ESO-1 gene was prepared by RT-PCR andsub-cloned into the prokaryotic expression vector pGEX-4T-2. The fusion protein, which expressed in E.coli BL21 cells, was purified by GST affinity chromgraphy and subjected to immunize Balb/c mouse. A kind of highly reactive and specific antiserum was obtained after immunization for several months. RT-PCR and ELISA also investigated the distribution of NY-ESO-1 as well as some other CT antigens in different normal and tumor tissues.Results: A high level expression of NY-ESO-1/GST fusion protein with 43 KD molecular weight was obtained after IPTG induced in E.coli. The specific polyclonal...