Antibody Engineering of Recombinant Matricellular CCN1 Protein-Specific ScFv Immunotoxin for Cancer Targeting

• Main Authors: Huei-Yin Chen, 陳慧頴
• Other Authors: Shr-Jeng Leu
• Format: Others
• Language: zh-TW
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/39038623300216971745

碩士 === 國立陽明大學 === 醫學生物技術暨檢驗學系 === 104 === CCN1, a secreted and extracellular matrix-associated protein, regulates multiple cellular activities, including cell survival, apoptosis, wound healing, and tissue repair. It has been shown that CCN1 is involved tumorigenesis of human oral cancer. Expression of CCN1 is enriched in oral cancer stem-like cells (OS-SLC). We have developed a CCN1-specific monoclonal antibody (mAb), designated YM-1E. Via the use of YM-1E, OS-SLC could be sorted out as the cell surface CCN1-positive and -negative cells. Cancer stemness was largely contained in the CCN1-positive cell population. This indicates that labeling of CCN1, via the use of YM-1E, can allow for targeting cancer stemness. Antibody-based therapeutics are of growing significance in cancer therapy. Single chain variable domain fragments (scFv) derived from mAb are powerful tools in research and clinical settings. Following successful cloning of cDNA of the YM-1E hybridoma, we have constructed an immunoglobulin gene fusion of the variable heavy (VH) and variable light (VL) chains that are joined together by a flexible peptide linker. We then further engineered the YM1E scFv fusion with Pseudomonas exotoxin A-PE38 for making the CCN1-targeted recombinant immunotoxin. Takes together, we wish that application of the CCN1-specific immunotoxin may make contribution to the effective cancer treatment in the new future.