CFTR-mutation specific applications of CFTR-directed monoclonal antibodies

Abstract

Background: Over the last decade novel monoclonal CFTR-specific antibodies have been developed. We here present a paired analysis to detect wild-type and mutant CFTR using Western blot analysis, flow cytometry and confocal microscopy in several cellular expression systems.

Methods: The following CFTR-specific antibodies were used; 217, 432, 450, 570, 769, 596, 660, L12B4 and 24.1. Mutant CFTR was detected in HEK293 cells transiently expressing the mutations; G542X, R1162X, F508del, N1303K, G551D, R117H, A455E.

Results: The majority of these antibodies are suitable for most applications tested. Using immunofluorescence, some antibodies can better detect mutant forms of CFTR (F508del and N1303K by mAbs 596 and 769), or display lower aspecific detection by Western blot analysis (mAbs 432, 450, 769 and 596) or immunofluorescence (mAbs 432, 450, 570 and 769).

Conclusion: Optimal detection of CFTR by monoclonal antibodies depends on CFTR mutation and the specific research application.

Keywords: Confocal microscopy; Flow cytometry; Monoclonal antibodies; Mutant CFTR; Paired analysis; Western blot.