Isolation and characterization of two monoclonal antibodies that recognize remote epitopes on the cell-binding domain of human fibronectin.

Experimental Cell Research
M KatayamaK Sekiguchi

Abstract

Two monoclonal anti-fibronectin antibodies that inhibit fibronectin-mediated cell adhesion have been established and characterized. One antibody, FN12-8, inhibited attachment of rat kidney fibroblasts on the fibronectin-coated substrate in a concentration-dependent manner, attaining a maximal inhibition of greater than 85% at 850 micrograms/ml. Another antibody, FN30-8, caused about 70% inhibition at a concentration as low as 0.85 microgram/ml, although further increase of the antibody concentration did not significantly augment the inhibitory effect. Immunoblot analysis with defined proteolytic fragments revealed that both antibodies are directed to the cell-binding domain of fibronectin. The epitopes for these antibodies were further narrowed down using recombinant cell-binding fragments expressed in Escherichia coli. FN12-8 recognized the 11.5-kDa cell-binding fragment previously characterized by Pierschbacher et al. (1981, Cell 26, 259-267), suggesting that FN12-8 blocks the Arg-Gly-Asp (RGD) cell adhesion signal. FN30-8 could not bind this fragment but did recognize a longer cell-binding fragment containing additional greater than 111 amino acid residues upstream of the 11.5-kDa fragment. Since the RGD-dependent cell adhes...Continue Reading

References

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Citations

Feb 15, 2005·Urology·Violeta MenéndezFrancisco García-López
Feb 1, 1995·Cell Adhesion and Communication·S K AkiyamaK M Yamada
May 1, 2009·Expert Opinion on Medical Diagnostics·Brian Keith McNeil, Robert H Getzenberg
Jan 25, 2005·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Shinya MizunoToshikazu Nakamura
Oct 4, 2003·The Journal of Biological Chemistry·Tomohiro KozakiKiyotoshi Sekiguchi
Jul 3, 1998·International Journal of Cancer. Journal International Du Cancer·S ThaonC Poustis-Delpont
Apr 12, 2017·Proceedings of the National Academy of Sciences of the United States of America·Alexandra F NiddamTara J Moriarty
Apr 24, 1999·Journal of the American Society of Nephrology : JASN·S WagaM Yokoyama

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