A sensitive enzyme immunoassay for the quantitation of human C5a/C5a(desArg) anaphylatoxin using a monoclonal antibody with specificity for a neoepitope

Complement Inflamm. 1991;8(1):13-24. doi: 10.1159/000463173.

Abstract

The direct quantitation of C5a/C5a(desArg) in human plasma was achieved by a specific and highly sensitive ELISA which is based on the neoepitope-specific monoclonal antibody C17/5. The error-prone removal of C5 from plasma prior to the assay is therefore not required. With a detection limit of 20 pg C5a/ml plasma, the sensitivity of this assay allowed to define normal ranges (1.94 +/- 1.49 ng C5a/nl, mean +/- SD) of C5a/C5a(desArg) in human blood plasma. This assay will also be applicable to the quantitation of C5a in specimens with low protein content where precipitation-based assays fail to accurately determine C5a. In addition, the mAb C17/5 turned out to efficiently block the binding of C5a/C5a (desArg) to its cellular receptor and therefore provides a valuable tool in the delineation of C5a effects in complex biologic systems in the presence of native C5, such as under in vivo conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / blood
  • Animals
  • Antibodies, Monoclonal*
  • Antibody Specificity / immunology
  • Blood Donors
  • Blood Platelets / metabolism
  • Blood Preservation
  • Calibration
  • Complement C5a / analysis*
  • Complement C5a / immunology
  • Complement C5a, des-Arginine / analysis*
  • Complement C5a, des-Arginine / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / analysis*
  • Guinea Pigs
  • Humans
  • In Vitro Techniques
  • Sensitivity and Specificity
  • Temperature

Substances

  • Antibodies, Monoclonal
  • Complement C5a, des-Arginine
  • Epitopes
  • Complement C5a
  • Adenosine Triphosphate