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. 1980 Dec 1;87(3):703–707. doi: 10.1083/jcb.87.3.703

Cell adhesion molecules: detection with univalent second antibody

PMCID: PMC2110804  PMID: 6970200

Abstract

Identification of cell surface molecules that play a role in cell-cell adhesion (here called cell adhesion molecules) has been achieved by demonstrating the inhibitory effect of univalent antibodies that bind these molecules in an in vitro assay of cell-cell adhesion. A more convenient reagent, intact (divalent) antibody, has been avoided because it might agglutinate the cells rather than blocking cell-cell adhesion. In this report, we show that intact rabbit immunoglobulin directed against certain cell surface molecules of Dictyostelium discoideum blocks cell-cell adhesion when the in vitro assay is performed in the presence of univalent goat anti-rabbit antibody. Under appropriate experimental conditions, the univalent second antibody blocks agglutination induced by the rabbit antibody without significantly interfering with its effect on cell-cell adhesion. This method promises to be useful for screening monoclonal antibodies raised against potential cell adhesion molecules because: (a) it allows for the screening of large numbers of antibody samples without preparation of univalent fragments; and (b) it requires much less antibody because of the greater affinity of divalent antibodies for antigens.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barondes S. H., Rosen S. D., Frazier W. A., Simpson D. L., Haywood P. L. Dictyostelium discoideum agglutinins (discoidins I and II). Methods Enzymol. 1978;50:306–312. doi: 10.1016/0076-6879(78)50033-5. [DOI] [PubMed] [Google Scholar]
  2. Beug H., Gerisch G., Kempff S., Riedel V., Cremer G. Specific inhibition of cell contact formation in Dictyostelium by univalent antibodies. Exp Cell Res. 1970 Nov;63(1):147–158. doi: 10.1016/0014-4827(70)90343-5. [DOI] [PubMed] [Google Scholar]
  3. Bozzaro S., Gerisch G. Contact sites in aggregating cells of Polysphondylium pallidum. J Mol Biol. 1978 Apr 5;120(2):265–279. doi: 10.1016/0022-2836(78)90067-0. [DOI] [PubMed] [Google Scholar]
  4. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  5. Geltosky J. E., Weseman J., Bakke A., Lerner R. A. Identification of a cell surface glycoprotein involved in cell aggregation in D. discoideum. Cell. 1979 Oct;18(2):391–398. doi: 10.1016/0092-8674(79)90058-8. [DOI] [PubMed] [Google Scholar]
  6. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  7. McDonough J. P., Springer W. R., Barondes S. H. Species-specific cell cohesion in cellular slime molds. Demonstration by several quantitative assays and with multiple species. Exp Cell Res. 1980 Jan;125(1):1–14. doi: 10.1016/0014-4827(80)90182-2. [DOI] [PubMed] [Google Scholar]
  8. Müller K., Gerisch G. A specific glycoprotein as the target site of adhesion blocking Fab in aggregating Dictyostelium cells. Nature. 1978 Aug 3;274(5670):445–449. doi: 10.1038/274445a0. [DOI] [PubMed] [Google Scholar]
  9. Ray J., Shinnick T., Lerner R. A mutation altering the function of a carbohydrate binding protein blocks cell-cell cohesion in developing Dictyostelium discoideum. Nature. 1979 May 17;279(5710):215–221. doi: 10.1038/279215a0. [DOI] [PubMed] [Google Scholar]
  10. Rosen S. D., Haywood P. L., Barondes S. H. Inhibition of intercellular adhesion in a cellular slime mould by univalent antibody against a cell-surface lectin. Nature. 1976 Sep 30;263(5576):425–427. doi: 10.1038/263425a0. [DOI] [PubMed] [Google Scholar]
  11. Springer W. R., Barondes S. H. Direct measurement of species-specific cohesion in cellular slime molds. J Cell Biol. 1978 Sep;78(3):937–942. doi: 10.1083/jcb.78.3.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Thiery J. P., Brackenbury R., Rutishauser U., Edelman G. M. Adhesion among neural cells of the chick embryo. II. Purification and characterization of a cell adhesion molecule from neural retina. J Biol Chem. 1977 Oct 10;252(19):6841–6845. [PubMed] [Google Scholar]

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