TY - JOUR
T1 - CFTR-mutation specific applications of CFTR-directed monoclonal antibodies
AU - van Meegen, M. A.
AU - Terheggen, S. W.J.
AU - Koymans, K. J.
AU - Vijftigschild, L. A.W.
AU - Dekkers, J. F.
AU - van der Ent, C. K.
AU - Beekman, J. M.
PY - 2013/9
Y1 - 2013/9
N2 - 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.
AB - 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.
KW - Confocal microscopy
KW - Flow cytometry
KW - Monoclonal antibodies
KW - Mutant CFTR
KW - Paired analysis
KW - Western blot
UR - http://www.scopus.com/inward/record.url?scp=84882831284&partnerID=8YFLogxK
U2 - 10.1016/j.jcf.2012.12.005
DO - 10.1016/j.jcf.2012.12.005
M3 - Article
C2 - 23317763
AN - SCOPUS:84882831284
SN - 1569-1993
VL - 12
SP - 487
EP - 496
JO - Journal of Cystic Fibrosis
JF - Journal of Cystic Fibrosis
IS - 5
ER -