Detection of anticentromere antibodies using recombinant human CENP-A protein.
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.


CONCLUSIONS
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.
Sun
D
D
Agouron Institute, La Jolla, California 92037-4696, USA.
Martinez
A
A
Sullivan
K F
KF
Sharp
G C
GC
Hoch
S O
SO
D000818:Animals; D001323:Autoantibodies / Q000032:analysis*; D001324:Autoantigens / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D002503:Centromere / Q000276:immunology*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005189:False Positive Reactions; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D007313:Insecta / Q000166:cytology / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D012595:Scleroderma, Systemic / Q000097:blood / Q000276:immunology
D001323:Autoantibodies; D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D011994:Recombinant Proteins