A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies

• Published in: PloS one Vol. 6; no. 7; p. e22533
• Main Authors: Carlring, Jennifer, De Leenheer, Evy, Heath, Andrew William
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 21.07.2011; Public Library of Science (PLoS)
• Subjects: Analysis; Animals; Antibodies, Bispecific - biosynthesis; Antigen-antibody reactions; Antigens; Antigens - immunology; B cells; Biochemistry - methods; Biology; Bispecific antibodies; Cancer; Chemistry; Cytometry; Dendritic cells; Drug dosages; Enzymes; Flow cytometry; Humans; Immunoassays; Immunoglobulin G; Immunoglobulins; Immunology; Infections; Infectious diseases; Lymphocytes; Lymphoma; Malnutrition; Medical research; Methods; Mice; Monoclonal antibodies; Oxidation-Reduction; Parenting; Pilot Projects; Proteins; Purification; Rats; Reproducibility of Results; Salts; Sodium; United Kingdom > UK
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0022533

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.