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Affinity Measurements by Band Shift and Competition ELISA

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Antibody Engineering

Part of the book series: Springer Lab Manuals ((SLM))

Abstract

The affinity constant (see Neri, Montigiani et al. 1996 for a simple introduction) is a useful parameter for the characterisation of antibody-antigen interactions. This constant can be seen as a “thermometre” which allows us to answer the following question: at certain conditions of concentration, to which extent does antibody A bind to its antigen B?

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References

  • Friguet B, Chaffotte AF, Djavadi-Ohaniance L, Goldberg ME (1985) Measurements of the true affinity constant in solution of antigen- antibody complexes by enzymelinked immunosorbent assay. J Immunol Methods 77: 305–319

    Article  PubMed  CAS  Google Scholar 

  • Montigiani S, Neri G, Neri P, Neri D (1996) Alanine substitutions in calmodulin-binding peptides result in unexpected affinity enhancement. J Mol Biol 258: 6–13

    Article  PubMed  CAS  Google Scholar 

  • Neri D, Montigiani S, Kirkham PM (1996) Biophysical methods for the determination of antibody-antigen affinities. Trends Biotechnol 14: 465–470

    Article  PubMed  CAS  Google Scholar 

  • Neri D, Petrul H, Winter G, Light Y, Marais R, Britton KE, Creighton AM (1996) Radioactive labeling of recombinant antibody fragments by phosphorylation using human casein kinase II and [gamma-32P]-ATP. Nat Biotechnol 14: 485–490

    Article  PubMed  CAS  Google Scholar 

  • Neri D, Prospero T, Petrul H, Winter G, Browne M, Vanderpant L (1996) Multipurpose high sensitivity luminescence analyzer (LUANA): use in gel electrophoresis. Biotechniques 20: 708–713

    PubMed  CAS  Google Scholar 

  • Nissim A, Hoogenboom HR, Tomlinson IM, Flynn G, Midgley C, Lane D, Winter G (1994) Antibody fragments from a ’single pot’ phage display library as immunochemical reagents. Embo J 13: 692–698

    PubMed  CAS  Google Scholar 

  • Viti F, Tarli L, Giovannoni L, Zardi L, Neri D (1999) Increased binding affinity and valence of recombinant antibody fragments lead to improved targeting of tumoral angiogenesis. Cancer Res 59: 347–352

    PubMed  CAS  Google Scholar 

  • Winter G, Griffiths AD, Hawkins RE, Hoogenboom HR (1994) Making antibodies by phage display technology. Annu Rev Immunol 12: 433–455

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Dario Neri .

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© 2001 Springer-Verlag Berlin Heidelberg

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Bumke, M.A., Neri, D. (2001). Affinity Measurements by Band Shift and Competition ELISA. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-04605-0_28

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  • DOI: https://doi.org/10.1007/978-3-662-04605-0_28

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-41354-7

  • Online ISBN: 978-3-662-04605-0

  • eBook Packages: Springer Book Archive

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