title: Microarray based on autodisplayed Ro proteins for medical diagnosis of systemic lupus erythematosus (SLE).
abstract: A microarray-based immunoassay for the detection of autoantibodies against Ro protein was developed using Escherichia coli with autodisplayed Ro proteins (Ro(+)-E. coli). Patient serum usually contains various antibodies against the outer membrane components of E. coli as well as autoantibodies against the Ro protein. Therefore, the conventional immunoassay based on Ro(+)-E. coli requires both wild type E. coli (blank test) and Ro(+)-E. coli, and both strains of E. coli must be prepared in situ for each individual test serum. In this study, we tested the feasibility of using several types of animal sera as a replacement for individual human sera. An immunoassay without the blank test was developed using Ro(+)-E. coli by (1) blocking with rabbit serum, and (2) cleaving the Fc region from antibodies using papain. Modified E. coli with autodisplayed Ro protein was immobilized to a surface-modified microplate and the applicability of the immunoassay without the blank test was demonstrated using sera from patients with systemic lupus erythematosus (SLE). Using this approach, a microarray-based fluorescence immunoassay with immobilized Ro(+)-E. coli was able to detect anti-Ro autoantibodies in SLE patient sera with high specificity and selectivity and improved efficiency.
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 134 Shin-chon-dong, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea.
authors.forename: Gu
authors.initials: G
authors.lastname: Yoo
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 134 Shin-chon-dong, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea.
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Bong
authors.affiliation: Department of Laboratory Medicine, College of Medicine, Yonsei University, Republic of Korea.
authors.forename: Sinyoung
authors.initials: S
authors.lastname: Kim
authors.affiliation: Institute of Pharmaceutical and Medical Chemistry, University of Muenster, Muenster, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Jose
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 134 Shin-chon-dong, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea. Electronic address: jcpyun@yonsei.ac.kr.
authors.forename: Jae-Chul
authors.initials: JC
authors.lastname: Pyun
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000032:analysis*; D015374:Biosensing Techniques / Q000379:methods*; D018914:Cells, Immobilized / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry; D015200:Fluoroimmunoassay / Q000379:methods; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000175:diagnosis*; D040081:Protein Array Analysis / Q000379:methods*; D011817:Rabbits; D012261:Ribonucleoproteins / Q000276:immunology*; D012680:Sensitivity and Specificity; D000099207:SS-A Antigen
chemical_list: D001323:Autoantibodies; D012261:Ribonucleoproteins; D000099207:SS-A Antigen
keywords: Autodisplay; Autoimmune; Fluorescence immunoassay; Microarray; Ro-protein; Systemic lupus erythematosus (SLE)