Abstract
International standards for anti-beta2 glycoprotein I (anti-β2GPI) testing are needed. We evaluated the suitability of polyclonal/monoclonal candidate reference materials (RM) for the assay. IgG/IgM anti-β2GPI were affinity-purified (AP) from high-positive antiphospholipid syndrome sera and IgG from HCAL clone supernatant. Igs were tested for purity by SDS-PAGE, pooled, concentrated, sterile-filtered and the protein concentration determined. One unit was defined as the binding activity of 1-μg/ml of AP anti-β2GPI Ig. IgG/IgM RM were each assigned a unit value using the respective AP material as a calibrator. Polyclonal/monoclonal RM and 30 samples were evaluated for linearity, unit equivalency and commutability. Polyclonal AP material was assigned a value of 100 U IgG and 15-U IgM anti-β2GPI, respectively. IgG-RM had a value of 270 IgG and the IgM-RM of 220.3 IgM anti-β2GPI U. The linearity (R2) of each RM curve for the various assays ranged from 0.96 to 0.99. Commutability samples fit very well within 95% prediction intervals and had excellent correlation when comparing assays. IgG and IgM polyclonal and IgG monoclonal RM displayed excellent linearity and commutability, being good candidates for better standardization of anti-β2GPI immunoassays.
| Original language | English |
|---|---|
| Pages (from-to) | 1317-1319 |
| Number of pages | 3 |
| Journal | Lupus |
| Volume | 23 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - Oct 8 2014 |
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Keywords
- Anti-β2glycoprotein I antibodies
- reference materials
- standardization
ASJC Scopus subject areas
- Rheumatology
- Medicine(all)
Cite this
International standards for IgG and IgM anti-β2glycoprotein antibody measurement. / Willis, R.; Grossi, C.; Orietta Borghi, M.; Martos-Sevilla, G.; Zegers, I.; Sheldon, J.; Meroni, P. L.
In: Lupus, Vol. 23, No. 12, 08.10.2014, p. 1317-1319.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - International standards for IgG and IgM anti-β2glycoprotein antibody measurement
AU - Willis, R.
AU - Grossi, C.
AU - Orietta Borghi, M.
AU - Martos-Sevilla, G.
AU - Zegers, I.
AU - Sheldon, J.
AU - Meroni, P. L.
PY - 2014/10/8
Y1 - 2014/10/8
N2 - International standards for anti-beta2 glycoprotein I (anti-β2GPI) testing are needed. We evaluated the suitability of polyclonal/monoclonal candidate reference materials (RM) for the assay. IgG/IgM anti-β2GPI were affinity-purified (AP) from high-positive antiphospholipid syndrome sera and IgG from HCAL clone supernatant. Igs were tested for purity by SDS-PAGE, pooled, concentrated, sterile-filtered and the protein concentration determined. One unit was defined as the binding activity of 1-μg/ml of AP anti-β2GPI Ig. IgG/IgM RM were each assigned a unit value using the respective AP material as a calibrator. Polyclonal/monoclonal RM and 30 samples were evaluated for linearity, unit equivalency and commutability. Polyclonal AP material was assigned a value of 100 U IgG and 15-U IgM anti-β2GPI, respectively. IgG-RM had a value of 270 IgG and the IgM-RM of 220.3 IgM anti-β2GPI U. The linearity (R2) of each RM curve for the various assays ranged from 0.96 to 0.99. Commutability samples fit very well within 95% prediction intervals and had excellent correlation when comparing assays. IgG and IgM polyclonal and IgG monoclonal RM displayed excellent linearity and commutability, being good candidates for better standardization of anti-β2GPI immunoassays.
AB - International standards for anti-beta2 glycoprotein I (anti-β2GPI) testing are needed. We evaluated the suitability of polyclonal/monoclonal candidate reference materials (RM) for the assay. IgG/IgM anti-β2GPI were affinity-purified (AP) from high-positive antiphospholipid syndrome sera and IgG from HCAL clone supernatant. Igs were tested for purity by SDS-PAGE, pooled, concentrated, sterile-filtered and the protein concentration determined. One unit was defined as the binding activity of 1-μg/ml of AP anti-β2GPI Ig. IgG/IgM RM were each assigned a unit value using the respective AP material as a calibrator. Polyclonal/monoclonal RM and 30 samples were evaluated for linearity, unit equivalency and commutability. Polyclonal AP material was assigned a value of 100 U IgG and 15-U IgM anti-β2GPI, respectively. IgG-RM had a value of 270 IgG and the IgM-RM of 220.3 IgM anti-β2GPI U. The linearity (R2) of each RM curve for the various assays ranged from 0.96 to 0.99. Commutability samples fit very well within 95% prediction intervals and had excellent correlation when comparing assays. IgG and IgM polyclonal and IgG monoclonal RM displayed excellent linearity and commutability, being good candidates for better standardization of anti-β2GPI immunoassays.
KW - Anti-β2glycoprotein I antibodies
KW - reference materials
KW - standardization
UR - http://www.scopus.com/inward/record.url?scp=84908673890&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84908673890&partnerID=8YFLogxK
U2 - 10.1177/0961203314544535
DO - 10.1177/0961203314544535
M3 - Article
C2 - 25228737
AN - SCOPUS:84908673890
VL - 23
SP - 1317
EP - 1319
JO - Lupus
JF - Lupus
SN - 0961-2033
IS - 12
ER -