The first quantitative comparison of immunohistochemical rabbit and mouse monoclonal antibody affinities using Biacore analysis

Here, we present the first quantitative investigation into the affinities of mouse and rabbit monoclonal antibodies for their antigens using Biacore(R) analysis. Our purpose was to determine the relationship between affinity and effectiveness for immunostaining. Our studies have focused on several important antibodies, including those to estrogen receptor, prouesterone receptor, CD3, and CD8. We Conclude that although antibody affinity can influence immunohistochemical intensity for some target epitopes, there is clearly a complex interplay between affinity, epitope availability, and specificity, which ultimately determines antibody performance. We have also shown that, in some instances, higher affinity antibodies are associated with "poorer" immunostaining performance. Antibody affinity is largely epitope dependent and varies depending on the target, and how it is presented to the host species immune system.