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).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
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.
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.
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.
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.
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.
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.
Mock, D. M. (1990) Sequential solid-phase assay for biotin based on 125I-labeled avidin. Methods Enzymol. 184, 224–233.
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.
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.
Vaidyanathan, G. and Zalutsky, M. R. (1990) Radioiodination of antibodies via N-succinimidyl 2,4-dimethoxy-3-(trialkylstannyl)benzoates. Bioconjugate Chem. 1, 387–393.
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.
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.
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.
Arnold Jr., L. J. (1985) Polylysine-drug conjugates. Methods Enzymol. 112, 270–285.
Pietersz, G. A. and McKenzie, I. F. (1992) Antibody conjugates for the treatment of cancer. Immunol. Rev. 129, 57–80.
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.
Dimroth, P. (1986) Preparation, characterization, and reconstitution of oxaloacetate decarboxylase from Klebsiella aerogenes, a sodium pump. Methods Enzymol. 125, 530–540.
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.
Kohanski, R. A. and Lane, D. (1990) Monovalent avidin affinity columns. Methods Enzymol. 184, 194–220.
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.
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.
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.
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.
Kronick, M. N. and Grossman, P. D. (1983) Immunoassay techniques with fluorescent phycobiliprotein conjugates. Clin. Chem. 29, 1582–1586.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights 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