Production and characterization of mouse monoclonal antibodies against human and mouse ribosomal proteins L5 and L11

master's thesis

• Author: Vedrana Jelenčić
• Mentor: Lidija Šver (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; Zagreb
• Defense date and country: 2010-07-02, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

The mammalian ribosome comprises 80 ribosomal proteins. Ribosomal proteins L5 and L11 are essential components of ribosome and recent evidence demonstrates that they participate in p53 tumor suppressor activation by interacting with Mdm2 and to bind to and influence activity of growth regulating transcription factor c-Myc. Due to lack of specific monoclonal antibodies against RpL5 and RpL11, which are required for further study of these and possible other extraribosomal functions of RpL11 and RpL5, we decided to produce mouse monoclonal antibodies against these ribosomal proteins. Ribosomal proteins are almost identical in mammals and therefore it is difficult to produce antibodies against them. Here we describe production of monoclonal antibodies directed against mouse and human RpL5 and RpL11 by immunizing mice with recombinant mouse Rps isolated from bacteria. Immune response against self-antigens in mice was successfully elicited by injecting large amounts of antigens combined with Freund's adjuvant. All produced mouse antibodies, including one against RpL11 and three against RpL5, could specifically recognize corresponding recombinant Rp on ELISA. While one monoclonal antibody against RpL5 could specifically recognize endogenous RpL5 in western blot analysis, other produced monoclonal antibodies require further testing. Beside to production of valuable antibodies for further characterization of RpL11 and RpL5 functions, described protocol show in general how to produce mouse monoclonal antibodies against highly conserved and thus less immunogenic antigens.

• Keywords: ribosomal proteins L5 and L11; p53 tumor suppressor; monoclonal antibodies
• Language: english
• URN:NBN: urn:nbn:hr:159:711272
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-06-02 11:38:10