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Detection of anticentromere antibodies using recombinant human CENP-A protein
D Sun1, A Martinez, K F Sullivan
1Agouron Institute, La Jolla, California 92037-4696, USA.
Objective:
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 = 100), 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.