Skip to main content

Preparation of Avidin Conjugates

  • Protocol
Book cover The Protein Protocols Handbook

Part of the book series: Springer Protocols Handbooks ((SPH))

  • 114 Accesses

Abstract

The high-affinity avidin-biotin system has found applications in different fields of biotechnology, including immunoassays, histochemistry, affinity chromatography, and drug delivery, to name a few. A brief description of avidin and avidin-like molecules, streptavidin, deglycosylated avidin, and NeutraLite avidin, is presented in the previous chapter (Chapter 46). With four biotin binding sites per molecule, the avidin family of proteins is capable of forming tight complexes with one or more biotinylated compounds (1). Typically, the avidin-biotin system is used to prepare signal-amplifying “sandwich” complexes between specificity reagents (e.g., antibodies) and detection reagents (e.g., fluorophores, enzymes, and so on). The specificity and detection reagents are independently conjugated, one with avidin and the other with biotin, or both with biotin, providing synthetic flexibility (2).

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 99.00
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. Green, N. M. (1975) Avidin, in Advances in Protein Chemistry, vol. 29 (Anfinsen, C. M., Edsall, J. T., and Richards, F. M., eds.), Academic, New York, 85–133.

    Google Scholar 

  2. Bayer, E. A. and Wilchek, M. (1980) The use of the avidin-biotin complex as a tool in molecular biology. Meth. Biochem. Anal. 26, 1–45.

    Article  CAS  Google Scholar 

  3. Niemeyer, C. M., Sano, T., Smith, C. L., and Cantor, C. R. (1994) Oligonucleotide-directed self-assembly of proteins: semisynthetic DNA-streptavidin hybrid molecules as connectors for the generation of macroscopic arrays and the construction of supramolecular bioconjugates. Nucleic Acids Res. 22, 5330–5339.

    Article  Google Scholar 

  4. Paganelli, G., Belloni, C., Magnani, P., Zito, F., Pasini, A., Sassi, I., Meroni, M., Mariani, M., Vignali, M., Siccardi, A. G., and Fazio, F. (1992) Two-step tumor targetting in ovarian cancer patients using biotinylated monoclonal antibodies and radioactive streptavidin. Eur. J. Nucl. Med. 19, 322–329.

    Article  PubMed  CAS  Google Scholar 

  5. van Osdol, W. W., Sung, C., Dedrick, R. L., and Weinstein, J. N. (1993) A distributed pharmacokinetic model of two-step imaging and treatment protocols: application to streptavidin-conjugated monoclonal antibodies and radiolabeled biotin. J. Nucl. Med. 34, 1552–1564.

    PubMed  Google Scholar 

  6. Markwell, M. A. K. (1982) A new solid-state reagent to iodinate proteins. I. Conditions for the efficient labeling of antiserum. Anal. Biochem. 125, 427–432.

    Article  PubMed  CAS  Google Scholar 

  7. Salacinski, P. R. P., McLean, C., Sykes, J. E. C., Clement-Jones, V. V., and Lowry, P. J. (1981) Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6,-tetrachloro-3α,6α-diphenyl glycoluril (iodogen). Anal. Biochem. 117, 136–146.

    Article  PubMed  CAS  Google Scholar 

  8. Mock, D. M. (1990) Sequential solid-phase assay for biotin based on 125I-labeled avidin. Methods Enzymol. 184, 224–233.

    Article  PubMed  CAS  Google Scholar 

  9. Bolton, A. E. and Hunter, W. M. (1973) The labeling of proteins to high specific radioactivity by conjugation to an 125I-containing acylating agent. Applications to the radioimmunoassay. Biochem. J. 133, 529–539.

    PubMed  CAS  Google Scholar 

  10. Vaidyanathan, G., Affleck, D. J., and Zalutsky, M. R. (1993) Radioiodination of proteins using N-succinimidyl 4-hydroxy-3-iodobenzoate. Bioconjugate Chem. 4, 78–84.

    Article  CAS  Google Scholar 

  11. Vaidyanathan, G. and Zalutsky, M. R. (1990) Radioiodination of antibodies via N-succinimidyl 2,4-dimethoxy-3-(trialkylstannyl)benzoates. Bioconjugate Chem. 1, 387–393.

    Article  CAS  Google Scholar 

  12. Hylarides, M. D., Wilbur, D. S., Reed, M. W., Hadley, S. W., Schroeder, J. R., and Grant, L. M. (1991) Preparation and in vivo evaluation of an N-(p-[125I]iodophenethyl)maleimide-antibody conjugate. Bioconjugate Chem. 2, 435–440.

    Article  CAS  Google Scholar 

  13. Arano, Y., Wakisaka, K., Ohmomo, Y., Uezono, T., Mukai, T., Motonari, H., Shiono, H., Sakahara, H., Konishi, J., Tanaka, C., and Yokoyama, A. (1994) Maleimidoethyl 3-(tri-n-butylstannyl)hippurate: A useful radioiodination reagent for protein radiopharmaceuticals to enhance target selective radioactivity localization. J. Med. Chem. 37, 2609–2618.

    Article  PubMed  CAS  Google Scholar 

  14. Willner, D., Trail, P. A., Hofstead, S. J., Dalton King, H., Lasch, S. J., Braslawsky, G. R., Greenfield, R. S., Kaneko, T., and Firestone, R. A. (1993) (6-Maleimidocaproyl)hydrazone of doxorubicin—A new derivative for the preparation of immunoconjugates of doxorubicin. Bioconjugate Chem. 4, 521–527.

    Article  CAS  Google Scholar 

  15. Arnold Jr., L. J. (1985) Polylysine-drug conjugates. Methods Enzymol. 112, 270–285.

    Article  PubMed  CAS  Google Scholar 

  16. Pietersz, G. A. and McKenzie, I. F. (1992) Antibody conjugates for the treatment of cancer. Immunol. Rev. 129, 57–80.

    Article  PubMed  CAS  Google Scholar 

  17. Orr, G. A. (1981) The use of the 2-iminobiotin-avidin interaction for the selective retrieval of labeled plasma membrane components. J. Biol. Chem. 256, 761–766.

    PubMed  CAS  Google Scholar 

  18. Dimroth, P. (1986) Preparation, characterization, and reconstitution of oxaloacetate decarboxylase from Klebsiella aerogenes, a sodium pump. Methods Enzymol. 125, 530–540.

    Article  PubMed  CAS  Google Scholar 

  19. Dean, P. D. G., Johnson, W. S., and Middle, F. S. (1985) Activation procedures, in Affinity Chromatography. A Practical Approach, IRL, Washington, DC, pp. 34,35.

    Google Scholar 

  20. Kohanski, R. A. and Lane, D. (1990) Monovalent avidin affinity columns. Methods Enzymol. 184, 194–220.

    Article  PubMed  CAS  Google Scholar 

  21. Chilkoti, A., Schwartz, B. L., Smith, R. D., Long, C. J., and Stayton, P. S. (1995) Engineered chimeric streptavidin tetramers as novel tools for bioseparations and drug delivery. Bio/Technology 13, 1198–1204.

    Article  PubMed  CAS  Google Scholar 

  22. Wong, S. S. (1991) Reactive groups of proteins and their modifying agents, in Chemistry of Protein Conjugation and Crosslinking, CRC, Boston, MA, pp. 7–48.

    Google Scholar 

  23. Carlsson, J., Drevin, H., and Axen, R. (1978) Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem. J. 173, 723–737.

    PubMed  CAS  Google Scholar 

  24. Haugland, R. P. (1995) Coupling of monoclonal antibodies with fluorophores. In Methods in Molecular Biology, vol. 45 (Davis, W. C., ed.), Humana, Totowa, NJ, pp. 205–221.

    Google Scholar 

  25. Kronick, M. N. and Grossman, P. D. (1983) Immunoassay techniques with fluorescent phycobiliprotein conjugates. Clin. Chem. 29, 1582–1586.

    PubMed  CAS  Google Scholar 

  26. Bongartz, J.-P., Aubertin, A.-M., Milhaud, P. G., and Lebleu, B. (1994) Improved biological activity of antisense oligonucleotides conjugated to a fusogenic peptide. Nucleic Acids Res. 22, 4681–4688.

    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

© 1996 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Haugland, R.P., Bhalgat, M.K. (1996). Preparation of Avidin Conjugates. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-60327-259-9_47

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-259-9_47

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-338-2

  • Online ISBN: 978-1-60327-259-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics