- 作者：Yoshinao Muro ; Kazumitsu Sugiura ; Akira Shiraki ; Norito Ishii ; Takashi Hashimoto ; Masashi Akiyama
- 关键词：PNP ; paraneoplastic pemphigus ; IPP ; immunoprecipitation ; DM ; dermatomyositis ; IPF ; idiopathic pulmonary fibrosis ; ACTD ; autoimmune connective tissue disease ; ILD ; interstitial lung diseases ; TnT ; transcription and translation ; T-PBS ; PBS containing 0.05% Tween20 ; RLU ; relative luminescence unit
- 刊名：Clinica Chimica Acta
- 出版年：15 February, 2014
- 年：2014
- 卷：429
- 期：Complete
- 页码：14-17
- 全文大小：514 K
Background
Paraneoplastic pemphigus (PNP) serum preferentially reacts with periplakin and envoplakin, which are plakin family proteins localized to desmosomes and intermediate filaments. Recently, anti-periplakin antibodies were also detected in patients with idiopathic pulmonary fibrosis (IPF). Although previous epitope-mapping studies showed multiple epitopes in each protein, enzyme-linked immunosorbent assays have used several truncated, but not full-length, recombinant proteins.Methods
This study aimed to produce full-length biotinylated recombinant proteins of periplakin and envoplakin for detection of autoantibodies by immunoprecipitation and ELISA. Serum from a PNP patient who had been confirmed as carrying anti-periplakin and anti-envoplakin antibodies in our previous study was used as a positive control. Sera from 15 patients with IPF were analyzed for both antibodies by immunoprecipitation and by ELISA.
Results
The PNP serum reacted strongly with the full-length recombinant proteins in immunoprecipitation and ELISA. Longitudinal serum samples from the PNP patient showed a clear decline of autoantibodies to both periplakin and envoplakin. None of the IPF sera showed both autoantibodies.
Conclusions
We found that the detection of anti-periplakin and anti-envoplakin antibodies using full-length recombinant proteins is useful immunoprecipitation and ELISA.