April 2009
Volume 50, Issue 13
ARVO Annual Meeting Abstract  |   April 2009
Antibody Mapping of Complement C3 Binding to CEP-BSA
Author Affiliations & Notes
  • L. A. Kuttner-Kondo
    Ophthalmic Research, Cleveland Clinic Foundation, Cleveland, Ohio
  • L. Hong
    Chemistry, Case Western Reserve University, Cleveland, Ohio
  • R. G. Salomon
    Chemistry, Case Western Reserve University, Cleveland, Ohio
  • J. G. Hollyfield
    Ophthalmic Research, Cleveland Clinic Foundation, Cleveland, Ohio
  • Footnotes
    Commercial Relationships  L.A. Kuttner-Kondo, None; L. Hong, None; R.G. Salomon, None; J.G. Hollyfield, None.
  • Footnotes
    Support  NIH Grants EY014240 and GM021249, NIH Infrastructure Grant EY015638, The Foundation Fighting Blindness, a Research to Prevent Blindness Challenge Grant, and State of Ohio - BRTT Grant
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 4942. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      L. A. Kuttner-Kondo, L. Hong, R. G. Salomon, J. G. Hollyfield; Antibody Mapping of Complement C3 Binding to CEP-BSA. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4942.

      Download citation file:

      © ARVO (1962-2015); The Authors (2016-present)

  • Supplements

Purpose: : Carboxyethyl pyrrole (CEP), an oxidation product of docosahexaenoic acid that is found adducted to proteins in drusen and plasma, may directly activate the alternative pathway of complement. This study was undertaken to localize the binding site of CEP adducted bovine serum albumin (CEP-BSA) on complement component C3 and begin an examination of the nature of the binding using the methyl ester of CEP-BSA for comparison.

Methods: : Sham-adducted BSA (BSA) or CEP-BSA was coated on ELISA plates. Subsequently, C3 fragments C3b, iC3b, C3c and/or C3d were added in equimolar quantities. Binding was assessed using a panel of anti-C3 monoclonal antibodies (mAbs) with specificity for C3c, C3d or neo-antigens.

Results: : An anti-C3c mAb clearly demonstrated the ability of C3b and iC3b to bind CEP-BSA. MAbs that recognize epitopes in C3d showed that C3d also binds CEP-BSA. In contrast, binding of these C3 fragments to BSA appears to be minimal. Using one anti-C3d mAb, C3d incubated on a Medisorp surface coated with either CEP-BSA or a methyl ester of CEP-BSA was detected at similar levels.

Conclusions: : These findings provide a basis for investigating a potential role of CEP-adducted protein in alternative pathway complement activation. As the methyl ester removes the carboxyl group, this opens up a possible role for the pyrrole ring in binding.

Keywords: oxidation/oxidative or free radical damage • age-related macular degeneration • inflammation 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.