Journal Article


Martin Clynes
Susan M Kennedy
Elizabeth Moran
Annemarie Larkin



pathology biosynthesis blotting western hl 60 cells antigens neoplasm cell line tumor breast neoplasms spleen humans antibodies monoclonal tissue fixation keratins immunohistochemistry neoplasm invasiveness immunology female chemistry fluorescent antibody technique genetics

Monoclonal antibody 5C3 raised against formalin fixed paraffin-embedded invasive breast tumour tissue: characterisation of its reactive antigen via immunoprecipitation and internal sequencing. (2005)

Abstract Monoclonal antibodies (MAbs) provide a powerful tool for the identification of novel tumour associated antigens. In an attempt to identify such an antigen, MAbs were generated by immunization with paraffin wax-embedded formalin-fixed invasive ductal breast tumour tissue from a patient who relapsed following an initial response to adjuvant chemotherapy. Extensive immunocytochemical and Western blot analysis of a range of cell lines and tissues including a series of pre- and post-chemotherapy treated invasive ductal breast carcinomas, with one of these MAbs, antibody 5C3, indicated that the 5C3 reactive antigen displayed a wide spectrum of reactivity amongst various human tumours. A reduced level of 5C3 expression was observed in non-cancerous archival breast tissues and breast cell lines and normal murine tissues compared to the expression observed in infiltrating breast tumour cells. Immunoprecipitation studies using the human ductal breast carcinoma cell line, ZR-75-1 resulted in the isolation of a 175 kDa reactive band which was excised from an SDS-PAGE gel and subjected to internal sequencing. Sequencing analysis and database searching revealed that this 175 kDa band represented a cytokeratin heteropolymer, composed of type I cytokeratin 9 and type II cytokeratin 6. Further studies confirmed that antibody 5C3 recognised this heteropolymer of cytokeratin 9 and 6 but not the individual cytokeratins. This novel method of MAb generation may facilitate the isolation of further potentially interesting cellular antigens. Characterisation of these novel antigens may identify specific disease targets with possible prognostic or predictive significance.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Martin Clynes, Susan M Kennedy, Elizabeth Moran, Annemarie Larkin

Experts in our system

Martin Clynes
Dublin City University
Total Publications: 209
Susan Kennedy
Dublin City University
Total Publications: 37
Annemarie Larkin
Dublin City University
Total Publications: 25