Skip to main content

Mini-Antibody Affinity Chromatography of Lysozyme

  • Protocol
Affinity Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 147))

  • 1424 Accesses

Abstract

Multiple noncovalent forces do play a role in the binding between a protein antigen and an antibody directed against that protein. An antigen is bound by the antigen binding site of an antibody that consists of three hypervariable segments (complementarity determining regions, CDRs) of the light chain (L1, L2, L3) and three of the heavy chain (H1, H2, H3) (see Fig. 1).

Schematic representation of the antigen-binding site. The hypervariable segment L1, L2, L3 of the light chain and H1, H2, H3 of the heavy chain of an immunoglobulin G molecule do have relatively small (e.g., L2) and relatively large (e.g., H1) contact areas with the epitope of an antigen (Ag).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 54.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Lerner R. A. (1982) Tapping the immunological repertoire to produce antibodies of predetermined specificity. Nature (London) 299, 592–596.

    Article  CAS  Google Scholar 

  2. Darsley M. J. and Rees A. R. (1985) Three distinct epitopes within the loop region of hen egg lysozyme defined with monoclonal antibodies. EMBO J. 4, 383–392.

    PubMed  CAS  Google Scholar 

  3. de la Paz P., Sutton B. J., Darsley M. J., and Rees A. R. (1986) Modeling of the combining sites of three anti-lysozyme monoclonal antibodies and of the complex between one of the antibodies and its epitope. EMBO J. 5, 415–425.

    PubMed  Google Scholar 

  4. Amitt A. G., Mariuzza R. A., Phillips S. E. V., and Poljak R. J. (1986) Three-dimensional structure of an antigen-antibody complex at 2.8 Å resolution. Science 233, 747–753.

    Article  Google Scholar 

  5. Fischmann T. O., Bentley G. A., Bhat T. N., Boulot G., Mariuzza R. A., Phillips S. E. V., Tello D., and Poljak R. J. (1991) Crystallographic refinement of the three-dimensional structure of the Fab D1.3-lysozyme complex at 2.5 Å resolution. J. Biol. Chem. 266, 12,915–12,920.

    PubMed  CAS  Google Scholar 

  6. Welling G. W., Geurts T., van Gorkum J., Damhof R. A., Drijfhout J. W., Bloemhoff W., and Welling-Wester S. (1990) Synthetic antibody fragment as ligand in immunoaffinity chromatography. J. Chromatogr. 512, 337–343.

    Article  PubMed  CAS  Google Scholar 

  7. Welling G. W., van Gorkum J., Damhof R. A., Drijfhout J. W., Bloemhoff W., and Welling-Wester S. (1991) A ten-residue fragment of an antibody (mini-antibody) directed against lysozyme as ligand in immunoaffinity chromatography. J. Chromatogr. 548, 235–242.

    Article  PubMed  CAS  Google Scholar 

  8. Lasonder E., Bloemhoff W., and Welling G.W. (1994) Interaction of lysozyme with synthetic anti-lysozyme D1.3 antibody fragments studied by affinity chromatography and surface plasmon resonance. J. Chromatogr. 676, 91–98.

    Article  CAS  Google Scholar 

  9. Ohlson S., Lundblad A., and Zopf D. (1988) Novel approach to affinity chromatography using “weak” monoclonal antibodies. Anal. Biochem., 169, 204–208.

    Article  PubMed  CAS  Google Scholar 

  10. Berry M. J, and Davies J. (1992) Use of antibody fragments in immunoaffinity chromatography. Comparison of FV fragments, VH fragments and paralog peptides. J. Chromatogr. 597, 239–245.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Humana Press Inc.

About this protocol

Cite this protocol

Welling, G.W., Welling-Wester, S. (2000). Mini-Antibody Affinity Chromatography of Lysozyme. In: Bailon, P., Ehrlich, G.K., Fung, WJ., Berthold, W. (eds) Affinity Chromatography. Methods in Molecular Biology, vol 147. Humana Press. https://doi.org/10.1007/978-1-60327-261-2_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-261-2_8

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-694-9

  • Online ISBN: 978-1-60327-261-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics