Problems and pitfalls with measurement of antibody affinity using solid phase binding in the ELISA.

@article{Underwood1993ProblemsAP,
  title={Problems and pitfalls with measurement of antibody affinity using solid phase binding in the ELISA.},
  author={Philippa Underwood},
  journal={Journal of immunological methods},
  year={1993},
  volume={164 1},
  pages={119-30}
}
Current methods of estimation of antibody affinity constants using ELISA assume homogeneous binding of antibody to the solid phase, despite many reports in the literature that this is not true. I have derived theoretical antibody binding curves for solid phase antigen assuming homogeneous antibody binding. I have compared these curves with a set of experimental binding curves of monoclonal antibodies to the serum protein fibronectin. The results conclusively show that while some monoclonal… CONTINUE READING