Using mimicry epitopes from phage displayed peptide library as substitute antigen for cyclophilin A in ELISA for detection of anti-CyPA autoantibody
- VernacularTitle:噬菌体模拟抗原替代亲环素A在测定自身抗体中的应用
- Author:
Fuyin LUO
;
Fangqiu LI
- Publication Type:Journal Article
- Keywords:
Bacteriophages;
Peptide library;
Lupus erythematosus,systemic;
Autoantibodies
- From:
Chinese Journal of Laboratory Medicine
2003;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To use mimicry epitopes from phage displayed peptide library as substitute for recombinant human cyclophilin A (rhCyPA) in detection of CyPA autoantibody in sera of patients with systemic lupus erythematosus (SLE).Methods Monoclonal antibody D4 and E6 were used as the capture antibody; the Ph.D.-12 p hage display peptide library was screened by biopanning. Ten phage clones with h igh affinity for McAb D4 and E6 (five for each) were chosen. Their inserts were sequenced. The amino sequence of mimicry epitope was subsequently reasoned out. Selected 12 peptide and previously obtained 7 peptide mimicry epitopes were used as coated antigen and compared with rhCyPA in ELISA for anti-CyPA autoantibody .Results Nine of ten phage clones with high affinity for McAb D4 and E6 (five for D4, four for E6) shared a consensus amino sequence HW EYTTSPHPRL. In comparative study of ELISA, 56 serum samples from SLE patients we re tested for anti-CyPA autoantibody, 28 samples were positive (50%) with 12 pe ptide mimicry antigen and 24 were positive (43%) with rhCyPA, 4 of 34 (11%) samp les from those with other autoimmune disorders were positive with both antigens, the coincidence rate between two methods was 95 6%. The research also showed t hat 12 peptide mimicry antigen has the advantages of rhCyPA and 7 peptide mimicr y antigen in detecting anti-CyPA autoantibody. Conclusions The specific 12 peptide screened from phage-displayed peptide library by ant i-CyPA monoclonal antibody can mimic epitope information of cyclophilin A well and be used as substitute for rhCyPA in ELISA.
