USE OF ANTIBODY ENGINEERING TO IMPROVE THE PROPERTIES OF ANTIBODIES AS A REAGENTS

Abstract

Two different antibody engineering strategies were explored to obtain a best-in-class antibody for use against mouse p53 with particular focus on the immunohistochemical (?IHC?) staining of paraffin-embedded mouse tissue sections. The first approach utilised a novel selection platform ?SBP-display? to engineer a known anti-human p53 antibody (DO-1) to bind the homologous epitope in mouse p53 which differs by a single amino acid residue. In the second approach, a known anti-mouse p53 antibody (PAb242) was engineered with the variable fragment (?Fv?) region grafted onto a human IgG1 scaffold. The resultant chimeric antibody (MH242) retained both the affinity and specificity of the parental antibody in IHC, ELISA, and Western Blot applications. IHC staining of mouse tissue sections confirmed that MH242 was able to detect mouse p53 protein with almost-zero background as compared with PAb242. Wide-spread adoption of such ?digitised? antibodies (with defined Fv sequences) should reduce experimental disparities that can commonly arise through variations in antibody quality.