DETECTION OF ANTICENTROMERE ANTIBODIES USING RECOMBINANT HUMAN CENP-APROTEIN

Objective. To evaluate CENP-A reactivity with anticentromere antibodie s (ACA) using recombinant protein (rCENP-A). Methods. Human CENP-A ant igen was overexpressed in insect cells using the baculovirus system. W e tested for ACA activity against the full-length recombinant polypept ide by immunoblot and by enzyme-linked immunosorbent assay (ELISA). Re sults. Of the ACA+ sera studied (n = 38), 95% were positive when teste d against the rCENP-A in the ELISA system. Of the ACA- sera (n = 100), only 2% gave false-positive results in the assay. There was good corr elation between the recombinant and bona fide antigens in assaying for ACA reactivity. Conclusion. CENP-A is a significant ACA target. The a vailability of the rCENP-A assay is a valuable adjunct to the previous ly described rCENP-B assay in analyses of the clinical significance of ACA.