Monoclonal Antibodies 1G12 and 6A12 to the N-domain of human angiotensin-converting enzyme: fine epitope mapping and antibody-based detection of ACE inhibitors in human blood.
Angiotensin I-converting enzyme (ACE), a key enzyme in cardiovascular pathophysiology, consists of two homologous domains (N- and C-), each bearing a Zn-dependent active site. ACE inhibitors are among the most prescribed drugs in the treatment of hypertension and cardiac failure. Fine epitope mapping of two monoclonal antibodies (mAb), 1G12 and 6A12, against the N-domain of human ACE, was developed using the N-domain 3D-structure and 21 single and double N-domain mutants. The binding of both mAbs to their epitopes on the N-domain of ACE is significantly diminished by the presence of the C-domain in the two-domain somatic tissue ACE and further diminished by the presence of sialic acid residues on the surface of blood ACE. The binding of these mAbs to blood ACE, however, increased dramatically (5-10-fold) in the presence of ACE inhibitors or EDTA, whereas the effect of these compounds on the binding of the mAbs to somatic tissue ACE was less pronounced and even less for truncated N-domain. This implies that the binding of ACE inhibitors or removal of Zn2+ from ACE active centers causes conformational adjustments in the mutual arrangement of N- and C-domains in the two-domain ACE molecule. As a result, the regions of the epitopes for mAb 1G12 and 6A12 on the N-domain, shielded in somatic ACE by the C-domain globule and additionally shielded in blood ACE by sialic acid residues in the oligosaccharide chains localized on Asn289 and Asn416, become unmasked. Therefore, we demonstrated a possibility to employ these mAbs (1G12 or 6A12) for detection and quantification of the presence of ACE inhibitors in human blood. This method should find wide application in monitoring clinical trials with ACE inhibitors as well as in the development of the approach for personalized medicine by these effective drugs.
Balyasnikova
Irina V
IV
Department of Anesthesiology, University of Illinois at Chicago, Illinois 60612, USA.
Skirgello
Olga E
OE
Binevski
Petr V
PV
Nesterovitch
Andrei B
AB
Albrecht
Ronald F
RF
Kost
Olga A
OA
Danilov
Sergei M
SM
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000097:blood* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001665:Binding Sites; D004492:Edetic Acid / Q000276:immunology; D018604:Epitope Mapping; D006801:Humans; D009154:Mutation; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011134:Polysaccharides / Q000276:immunology; D017434:Protein Structure, Tertiary; D015032:Zinc / Q000378:metabolism
C521204:1G12 monoclonal antibody; C521205:6A12 monoclonal antibody; D000806:Angiotensin-Converting Enzyme Inhibitors; D000911:Antibodies, Monoclonal; D011134:Polysaccharides; D004492:Edetic Acid; D007703:Peptidyl-Dipeptidase A; D015032:Zinc