CTL - Tumor Cell Interaction: The Generation of Molecular Probes Capable of Monitoring the HLA-A*0201-HER-2/neu Peptide Complex
Annual summary rept. 27 Feb 2005-26 Feb 2006
ROSWELL PARK CANCER INST BUFFALO NY
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The overall goal of this project is to probe the CTL - tumor cell interaction by generating scFv probes that are able to recognize the HLA- A0201-HER-2neu369-377 peptide complex. In the 12 month period covered by this report I aimed to improve the affinity of HLA-A0201- HER-2neu369-377 peptide complex-specific scFv 2.3.5 which was isolated during the first 12 month period of this project. Utilizing three rounds of site-directed mutagenesis in the VL CDR3 and VL CDR1 of scFv 2.3.5, I have generated and isolated a scFv clone named 2.3.5- 58-53 which exhibited markedly higher affinity for the HLA-A0201-HER-2neu369-377 peptide complex than the parental scFv 2.3.5. I will utilize scFv 2.3.5-58-53 to analyze the levels of HLA-A0201 HER-2neu369-377 complexes expressed by breast carcinoma cell lines and breast carcinoma lesions in order to determine if there are correlations between the levels of antigen processing machinery components HLA class I antigens Beta2-microglobulin andor HER-2neu on the levels of HLA-A0201 HER-2neu369-377 complex on breast carcinoma cells.
- Medicine and Medical Research