Advertisement

Characterization and Selection Criteria of Monoclonal Antibodies for Tumor Imaging Studies

  • A. A. Noujaim
  • B. M. Longenecker
  • M. R. Suresh
  • C. J. Turner
  • T. R. Sykes
  • G. D. MacLean
Conference paper
Part of the NATO ASI Series book series (NSSA, volume 152)

Abstract

Evidence now exists that abnormalities of cell-surface molecular architecture are often associated with cancer. These subtle, yet dramatic changes, are the result of alterations in the gene regulation mechanism leading to either a blockage, stimulation or neosynthesis of cell surface molecules which, under normal conditions, would mediate cell-cell interaction and cellular differentiation (1,2). Thus, cells which have been oncologically transformed either in vitro or in vivo, will display sufficiently different profiles from their progenitor cells that they can be targeted by means of monoclonal antibodies (MAbs) which in turn could carry a number of cytotoxic agents or radionuclides. On the surface, this approach appears to be not only elegant but rather simple. In actual fact, the problems encountered in the design of such probes are predicated by the availability of the right MAb which recognizes not only a particularly well-defined chemical structure, but also an epitope of such structure consistent with the cell membrane environment of the tumor cell. Even if such a “magic bullet” were to be found, other dynamic characteristics of cell membrane constituents alter, to an extent, the degree of specificity of the selected MAb. While fundamental questions are often raised as to the importance of marker turnover as a function of cell differentiation, others are related to the relevance of the animal models so often used prior to immunoscintigraphic studies. Such diversities frequently pose considerable difficulties when selecting the appropriate antibody for clinical trials. An excellent review of this subject has been presented by Larson (3). In our laboratory, we have been interested in generating MAbs against cancer-associated carbohydrate markers and the use of such MAbs for the in vivo and in vitro detection of human tumors (4,5). Some of our experience in selecting and characterizing these MAbs is described in this paper.

Keywords

Ammonium Sulfate Protein Recovery Detergent Extract Free Sulfhydryl Group Bifunctional Chelate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    S. Hakamori, and R. Kannagi, Glycosphingolipids as Tumor-Associated and Differentiation Markers, J.N.C.I. 71:231 (1983).Google Scholar
  2. 2.
    R.O. Hynes, and C.F. Fox, Tumor Cell Surface and Malignancy, Prog. Clin. Biol. Res. Monogr. 41:1 (1980).Google Scholar
  3. 3.
    S.M. Larson, J.A. Carrasquillo, and J. Reynolds, Radioimmunodetection and Radioimmunotherapy, Cancer Invest. 2:363 (1984).PubMedCrossRefGoogle Scholar
  4. 4.
    A. A. Noujaim, S. Selvaraj, M.R. Suresh, G.D. MacLean, D. Willans, C.J. Turner, D. Haines, and B.M. Longenecker, A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors, Nuklearmedizin 26:1 (1986).Google Scholar
  5. 5.
    T. Feizi, Demonstration by Monoclonal Antibodies that Carbohydrate Structures of Glycoproteins and Glycolipids are Neo-Developmental Antigens, Nature 314:53 (1985).PubMedCrossRefGoogle Scholar
  6. 6.
    A.A. Noujaim, A. Shysh, P. Zabel, J. Bray, M.R. Suresh, and B.M. Longenecker, Thomsen-Friedenreich Antigen: An Important Marker for the Radioimmunodetection of Cancer Using Molecular Probes, in: “Radioimmunoimaging and Radioimmunotherapy,” S.W. Burchiel, B.A. Rhodes, eds., Elsevier Publ., New York (1983).Google Scholar
  7. 7.
    S.E. Ritzman, ed. “Protein Abnormalities”, Vol. I, Alan R. Liss, Inc., New York (1982).Google Scholar
  8. 8.
    R. Lindmark, K. Thoren-Tolling, and J. Sjoquist, Binding of Immunoglobulins to Protein A and Immunoglobulins in Mammalian Sera, J. Immunol. Meth. 62:1 (1983).CrossRefGoogle Scholar
  9. 9.
    P.L. Ey, S.J. Prowse, and C.J. Jenkin, Isolation of Pure IgG, IgG-2a and IgG-2b Immunoglobulins from Mouse Serum using Protein A Sepharose, Immunochemistry 15:429 (1978).PubMedCrossRefGoogle Scholar
  10. 10.
    C.J. Van Oss, Isolation and Characterization of Immunoglobulins, Sep. Purif. Meth. 11;131 (1982).CrossRefGoogle Scholar
  11. 11.
    J. Steensgaard, and A.S. Johansen, Biochemical Aspects of Immune Complex Formation and Immune Complex Diseases. Allergy 35:457 (1980).PubMedCrossRefGoogle Scholar
  12. 12.
    M.W. Steward, and J. Steensgaard, “Antibody Affinity: Thermodynamic Aspects and Biological Significance,” CRC Press, Boca Raton, Fla. (1984).Google Scholar
  13. 13.
    J.R. Stephenson, J.M. Lee, and P.P. Willon-Smith, Production and Purification of Murine Monoclonal Antibodies: Aberrant Elution from Protein A Sepharose 4B, Anal. Biochem. 142:189 (1984).PubMedCrossRefGoogle Scholar
  14. 14.
    E. Regoeczi, “Iodine Labelled Plasma Proteins, “CRC Press, Boca Raton, Fla. (1984).Google Scholar
  15. 15.
    S.M. Yeh, D.G. Sherman, and C.F. Meares, A New Route to “Bifunctional” Chelating Agents: Conversion of Amino Acid Analogs to Ethylenediaminetetraacetic Acid, Anal. Biochem. 100:152 (1979).PubMedCrossRefGoogle Scholar
  16. 16.
    C.S. Leung, “The Covalent Attachment of ‘Bifunctional’ Chelates to Macromolecules and Their Use as Physical Probes in Biological Systems,” Ph.D. Thesis, Univ. Calif., Davis (1977).Google Scholar
  17. 17.
    L.H. DeReimer, C.F. Meares, D.A. Goodwin, and C.I. Diamanti, BLEDTA: Tumor Localization by a Bleomycin Analogue Containing a Metal-Chelating Group, J. Med. Chem. 22:1019 (1979).CrossRefGoogle Scholar
  18. 18.
    G.E. Means, and R.E. Feeney, “Chemical Modification of Proteins,” Holden-Day, San Francisco (1971).Google Scholar
  19. 19.
    C.F. Meares, M.J. McCall, D.T. Reardon, D.A. Goodwin, C.I. Diamanti, and M. McTigue, Conjugation of Antibodies with Bifunctional Chelating Agents: Isothiocyanate and Bromoacetamide Reagents, Methods of Analysis and Subsequent Addition of Metal Ions, Anal. Biochem. 142:68 (1984).PubMedCrossRefGoogle Scholar
  20. 20.
    S.M. Yeh, I. New Bifunctional Chelates as Biophysical Probes II. Diffusion Enhanced Lanthanide Energy Transfer Studies of Transferrin, Ph.D. Thesis, Univ. of Calif., Davis (1979).Google Scholar
  21. 21.
    C.J. Turner, T.R. Sykes, R.C. Gaudreault, B.M. Longenecker, and A.A. Noujaim, Determination of Metal Ion Impurities in Radiogallium Preparations and Their Effects on the Radiolabelling of Chelated Proteins, in: “Current Applications in Radiopharmacology,” M. W. Billinghurst, ed., Pergamon Press, Toronto (1986).Google Scholar
  22. 22.
    H.S. Penefsky, A Centrifuged-Column Procedure for the Measurement of Ligand Binding by Beef Heart F1, Meth. Enzymol. 56:527 (1979).PubMedCrossRefGoogle Scholar
  23. 23.
    A. Saul, M. Don, A Rapid Method of “Concentrating Proteins in Small Volumes with High Recovery Using Sephadex G-25, Anal. Biochem. 138:451 (1984).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1988

Authors and Affiliations

  • A. A. Noujaim
    • 1
  • B. M. Longenecker
    • 1
    • 2
  • M. R. Suresh
    • 1
    • 3
  • C. J. Turner
    • 1
  • T. R. Sykes
    • 1
    • 2
  • G. D. MacLean
    • 1
    • 4
  1. 1.Faculty of Pharmacy and Department of ImmunologyUniversity of AlbertaEdmontonCanada
  2. 2.Department of ImmunologyUniversity of AlbertaEdmontonCanada
  3. 3.Biomira, Inc.EdmontonCanada
  4. 4.Cross Cancer InstituteEdmontonCanada

Personalised recommendations