Epitope Mapping Studies of Snake Venom Phospholipase A2 Using Monoclonal Antibodies

  • Bradley G. Stiles
  • John L. Middlebrook
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 303)


Fifteen different monoclonal antibodies developed against cpseudexin, a snake venom phospholipase A2 with presynaptic cneurotoxicity, were screened for linear epitope recognition. cPeptides (9-mers) spanning pseudexin were synthesized by using calanine-derivatized polyethylene pins and subsequently probed with cantibody. Four antibodies bound to toxin peptides and were detected cwith an enzyme-linked immunosorbent assay. Three of the bound cantibodies recognized a site important in calcium binding and the cinterlocking of dimeric forms of snake venom phospholipase A2. cAnalogous regions from other phospholipases were synthesized and cprobed with the four reactive antibodies. A good correlation was cfound between the reactivity of whole molecule phospholipases and cpeptide regions with the antibodies. Monoclonal antibodies cneutralizing the lethal or enzymatic effects of pseudexin did not crecognize any linear epitopes.


Epitope Mapping Linear Epitope Enzyme Link Immuno Sorbent Assay Toxin Peptide Absorbance Means 
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  1. 1.
    Kini, R.M. and Evans, H. (1989) Toxicon 27, 613–635.PubMedCrossRefGoogle Scholar
  2. 2.
    Middlebrook, J. L. and Kaiser I. I. (1989) Toxicon 27, 965–977.PubMedCrossRefGoogle Scholar
  3. 3.
    Middlebrook, J. L. (1990) Toxicon, In Press.Google Scholar
  4. 4.
    Geysen, H.M., Meloen, R.H., and Barteling, S.J. (1984) Proc. Natl. Acad. Sci. USA 81, 3998–4002.PubMedCrossRefGoogle Scholar
  5. 5.
    Aird, S.D., Kaiser, I.I., Lewis, R.V. and Kruggel, W.G.(1985) Biochemistry 24, 7054–7058.PubMedCrossRefGoogle Scholar
  6. 6.
    Bieber, A.L., Becker, R.R., McParland, R., Hunt, D.F., Shabanowitz, J., Yates, J.R., Martino, P.A., and Johnson, G.R. (1990) Bioch. Biophys. Acta 1037, 413–421.CrossRefGoogle Scholar
  7. 7.
    Dufton, M.J. and Hider, R.C. (1983) Eur. J. Biochem. 137, 545–551.PubMedCrossRefGoogle Scholar
  8. 8.
    Mebs, D. (1985) List of Biologically Active Components from Snake Venoms. Pergamon Press, Oxford, England.Google Scholar
  9. 9.
    Schmidt, J.J., and Middlebrook, J.L (1989) Toxicon 27, 805–818.PubMedCrossRefGoogle Scholar
  10. 10.
    Keith, C., Feldman, D.S., Deganello, S., Glick, J., Ward, K.B., Jones, E.O., and Sigler, P.B. (1981) J. Biol. Chem. 256, 8602–8607.PubMedGoogle Scholar
  11. 11.
    Renetseder, R., Brunie, S., Dijkstra, B.W., Drenth, J., and Sigler, P.B. (1985) J. Biol. Chem. 260, 11627–11634.PubMedGoogle Scholar
  12. 12.
    Amit, A.G., Mariuzza, R.A., Philips, S.E.V., and Poljak, R.J. (1986) Science 233, 747–753.PubMedCrossRefGoogle Scholar
  13. 13.
    Padlan, E.A., Silverton, E.W., Sheriff, S., Cohen, G.H., Smith-Gill, S.J., and Davies, D.R. (1989) Proc. Natl. Acad. Sci. USA 86, 5938–5942.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Bradley G. Stiles
    • 1
  • John L. Middlebrook
    • 1
  1. 1.Department of Toxicology Pathophysiology DivisionU.S. Army Medical Research Institute of Infectious DiseasesFrederickUSA

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