A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies.
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 solu...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3141073?pdf=render |
| _version_ | 1830231719723139072 |
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| author | Jennifer Carlring Evy De Leenheer Andrew William Heath |
| author_facet | Jennifer Carlring Evy De Leenheer Andrew William Heath |
| author_sort | Jennifer Carlring |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-18T11:19:48Z |
| format | Article |
| id | doaj.art-6607d53e3c1e44dda470eaf339710ceb |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-18T11:19:48Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-6607d53e3c1e44dda470eaf339710ceb2022-12-21T21:09:50ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0167e2253310.1371/journal.pone.0022533A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies.Jennifer CarlringEvy De LeenheerAndrew William HeathWe 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.http://europepmc.org/articles/PMC3141073?pdf=render |
| spellingShingle | Jennifer Carlring Evy De Leenheer Andrew William Heath A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies. PLoS ONE |
| title | A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies. |
| title_full | A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies. |
| title_fullStr | A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies. |
| title_full_unstemmed | A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies. |
| title_short | A novel redox method for rapid production of functional bi-specific antibodies for use in early pilot studies. |
| title_sort | novel redox method for rapid production of functional bi specific antibodies for use in early pilot studies |
| url | http://europepmc.org/articles/PMC3141073?pdf=render |
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