상피세포암 마커 Epithin/PRSS14 에 대한 단일클론항체의 제작과 이용

Title
상피세포암 마커 Epithin/PRSS14 에 대한 단일클론항체의 제작과 이용
Authors
정한나
Keywords
상피세포암마커epithinprss14에대한단일클론항체의제작과이용
Issue Date
2011
Publisher
인하대학교
Abstract
Epithin/PRSS14 is a type Ⅱ transmembrane serine protease that is overexpresses in many epithelial cancers and activates many well known tumor associated proteins. In order to generate useful reagents, monoclonal antibodies (mAbs) against epithin/PRSS14, mAbs were generated by hybridoma technology using strategically designed antigenic peptides. Hybridoma clones with high antibody titer in Enzyme-Linked Immunosorbent Assay (ELISA) were further selected via effectivity in vitro in the functional assays. Four mAbs revealed inhibitory effect on cell migration of epithelial cancer cells as well as endothelial cells induced by Epithin/PRSS14. In addition, some of these antibodies showed specific binding to antigen present in mouse and/or human in flow cytometric assays. They also detected specific human proteins in western blotting. Generated monoclonal antibodies in this study revealed the great potentials for the use in the development of diagnosis and therapeutics agents specific for epithelial cancer.
Description
1. Introduction 1 2. Meterials and Methods 4 2.1 Antigenic peptide 4 2.2 DNA cloning 5 2.3 Cell culture and transfection 5 2.4 Epi-SP peptide Immunization and hybridoma fusion 6 2.5 ELISA - Indirect ELISA/sandwich ELISA 7 2.6 Wound healing migration assay 8 2.7 Flow cytometry analysis 8 2.8 Western blot analysis 9 3. Results 11 3.1 Generation and selection hybridoma against epithin/PRSS14 by fusion 11 3.2 mAbs inhibit the cell migration of both epithelial cancer cell and endothelial cell induced by epithin/PRSS14 12 3.3 mAbs bind to cell surface epithin/PRSS14 specifically: in flow cytometry 13 3.4 mAbs can detect human protein in western blot 14 3.5 Detection of soluble protein using sandwich ELISA 15 4. Discussion 16 5. Figures 19 6. Reference 34
URI
http://dspace.inha.ac.kr/handle/10505/22525
Appears in Collections:
College of Natural Science(자연과학대학) > Biological Sciences (생명과학) > Theses(생명과학 석박사 학위논문)
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