a novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies新颖的氧化还原方法快速生产功能性bi-specific抗体用于早期的试验研究.pdfVIP

A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies Jennifer Carlring*, Evy De Leenheer, Andrew William Heath Academic Unit of Immunology and Infectious Diseases, Department of Infection and Immunity, School of Medicine and Biomedical Sciences, University of Sheffield, Sheffield, United Kingdom 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. Citation: Carlring J, De Leenheer E, Heath AW (2011) A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies. PLoS ONE 6(7): e22533. doi:10.1371/journal.pone.0022533 Editor: K