Abstract
Despite the fact that a growing number of recombinant antibodies has been isolated from phage display libraries, still many antibody specificities are available from hybridoma cell lines. Here, a method is presented to obtain the genetic information for the antigen binding part of the antibody from hybridoma cells, and to assemble it into a functional bacterially expressed fusion protein (scFv fragment). To achieve this, vectors have been constructed which combine the two variable regions (Vh and Vl) with a peptide linker to yield an scFv fragment. The genetic information for Vh and Vl is amplified from hybridoma cells using the polymerase chain reaction (PCR) with antibody specific primers.
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© 2001 Springer-Verlag Berlin Heidelberg
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Breitling, F., Moosmayer, D., Brocks, B., Dübel, S. (2001). Construction of scFv from Hybridoma by Two-Step Cloning. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-04605-0_3
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DOI: https://doi.org/10.1007/978-3-662-04605-0_3
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-41354-7
Online ISBN: 978-3-662-04605-0
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