Detection of anticentromere antibodies using recombinant human CENP-A protein

✍ Scribed by Dongxu Sun; Antigona Martinez; Kevin F. Sullivan; Gordon C. Sharp; Sallie O. Hoch

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 503 KB
• Volume: 39
• Category: Article
• ISSN: 0004-3591
• DOI: 10.1002/art.1780390520

✦ Synopsis

To evaluate CENP-A reactivity with anticentromere antibodies (ACA) using recombinant protein (rCENP-A).

Methods. Human CENP-A antigen was overexpressed in insect cells using the baculovirus system. We tested for ACA activity against the full-length recombinant polypeptide by immunoblot and by enzyme-linked immunosorbent assay (ELISA).

Results. Of the ACA+ sera studied (n = 38), 95% were positive when tested against the rCENP-A in the ELISA system. Of the ACA-sera (n = 1001, only 2% gave false-positive results in the assay. There was good correlation between the recombinant and bona fide antigens in assaying for ACA reactivity.

Conclusion. CENP-A is a significant ACA target. The availability of the rCENP-A assay is a valuable adjunct to the previously described rCENP-B assay in analyses of the clinical significance of ACA.

Autoantibodies are characteristic of the rheumatic disease systemic sclerosis (SSc; scleroderma) (1,2). Anticentromere antibodies (ACA) are the primary specificity associated with the limited cutaneous form of the disease, which includes the CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasias). The molecular nature of a number of autoantigens has been