title: A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies.
abstract: We demonstrate here a rapid alternative method for the production of functional bi-specific antibodies using the mild reducing agent 2-mercaptoethanesulfonic acid sodium salt (MESNA). Following reduction of a mixture of two monoclonal antibodies with MESNA to break inter heavy chain bonds, this solution is dialysed under oxidising conditions and antibodies are allowed to reform. During this reaction a mixture of antibodies is formed, including parental antibodies and bi-specific antibody. Bi-specific antibodies are purified over two sequential affinity columns. Following purification, bi-specificity of antibodies is determined in enzyme-linked immunosorbent assays and by flow cytometry. Using this redox method we have been successful in producing hybrid and same-species bi-specific antibodies in a time frame of 6-10 working days, making this production method a time saving alternative to the time-consuming traditional heterohybridoma technology for the production of bi-specific antibodies for use in early pilot studies. The use of both rat and mouse IgG antibodies forming a rat/mouse bi-specific antibody as well as producing a pure mouse bi-specific antibody and a pure rat bi-specific antibody demonstrates the flexibility of this production method.
authors.affiliation: Academic Unit of Immunology and Infectious Diseases, Department of Infection and Immunity, School of Medicine and Biomedical Sciences, University of Sheffield, Sheffield, United Kingdom. j.u.carlring-wright@shef.ac.uk
authors.forename: Jennifer
authors.initials: J
authors.lastname: Carlring
authors.forename: Evy
authors.initials: E
authors.lastname: De Leenheer
authors.forename: Andrew William
authors.initials: AW
authors.lastname: Heath
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis*; D000941:Antigens / Q000276:immunology; D001671:Biochemistry / Q000379:methods*; D006801:Humans; D051379:Mice; D010084:Oxidation-Reduction; D010865:Pilot Projects; D051381:Rats; D015203:Reproducibility of Results
chemical_list: D018033:Antibodies, Bispecific; D000941:Antigens