PMID- 10594571
OWN - NLM
STAT- MEDLINE
DCOM- 20000113
LR  - 20190513
IS  - 0009-9104 (Print)
IS  - 1365-2249 (Electronic)
IS  - 0009-9104 (Linking)
VI  - 118
IP  - 3
DP  - 1999 Dec
TI  - Binding properties of antibodies to prothrombin and beta2-glycoprotein I 
      (beta2-GPI) assayed by ELISA and dot blot.
PG  - 480-6
AB  - Most anti-phospholipid antibodies (aPL) associated with the anti-phospholipid 
      syndrome are autoantibodies with specificity towards beta2-GPI (anti-beta2-GPI) 
      or prothrombin (anti-II). They are mainly screened by ELISA using polyoxygenated 
      plates. However, some authors have claimed that immunoblotting can also be used. 
      Exposure of cryptic epitopes or increase of antigen density on its binding to 
      either phospholipids or suitable plastic surfaces are the two hypotheses proposed 
      for the interaction of beta2-GPI or prothrombin with their antibodies. Forty-five 
      patients with aPL were studied: 25 with lupus anti-coagulant (LA) and 
      anti-cardiolipin antibodies (aCL), 10 with LA alone and 10 with aCL but negative 
      LA. All patients with LA and aCL were positive for anti-beta2-GPI by ELISA and 
      dot blot, while 15/25 had anti-IIELISA and 14 of them also had anti-II by dot 
      blot assay. No patient with LA alone tested positive for anti-beta2-GPI by ELISA 
      or dot blot, whereas 6/10 had anti-IIELISA (five of them were also positive by 
      dot blot). Four out of 10 aCL-positive patients had anti-beta2-GPI by ELISA and 
      dot blot, while none of this group had anti-II by ELISA or dot blot. Antibody 
      binding to beta2-GPI or prothrombin in both ELISA and dot blot was significantly 
      reduced by phospholipid liposomes mixed together with beta2-GPI or prothrombin, 
      whereas liposomal eluants retained it in both assays. Parallel fluid-phase 
      inhibition experiments using increasing concentrations (up to 200 microg/ml) of 
      beta2-GPI or prothrombin demonstrated that antibody binding reduction was more 
      evident on dot blot than on ELISA. It was almost completely abolished on dot 
      blot, while on ELISA a moderate inhibition was achieved even at the highest 
      protein concentration. However, antibody binding on ELISA was virtually abolished 
      when diluted sera were incubated with high protein concentrations applied to 
      nitrocellulose membranes. We could infer that ELISA and dot blot detect 
      antibodies with some differences in avidity but directed against native epitopes 
      on beta2-GPI and prothrombin.
FAU - Forastiero, R R
AU  - Forastiero RR
AD  - Favaloro University and Institute of Cardiology and Cardiovascular Surgery, 
      Favaloro Foundation, Buenos Aires, Argentina.
FAU - Martinuzzo, M E
AU  - Martinuzzo ME
FAU - Carreras, L O
AU  - Carreras LO
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Clin Exp Immunol
JT  - Clinical and experimental immunology
JID - 0057202
RN  - 0 (Antibodies, Anticardiolipin)
RN  - 0 (Antibodies, Antiphospholipid)
RN  - 0 (Anticoagulants)
RN  - 0 (Blood Proteins)
RN  - 0 (Glycoproteins)
RN  - 0 (Liposomes)
RN  - 0 (beta 2-Glycoprotein I)
RN  - 9001-26-7 (Prothrombin)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Antibodies, Anticardiolipin/metabolism
MH  - Antibodies, Antiphospholipid/*metabolism
MH  - Antibody Specificity/immunology
MH  - Anticoagulants/*immunology
MH  - Binding, Competitive/drug effects/immunology
MH  - Blood Proteins/immunology/pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Female
MH  - Glycoproteins/*immunology
MH  - Humans
MH  - Immunoblotting
MH  - Liposomes/pharmacology
MH  - Male
MH  - Middle Aged
MH  - Prothrombin/*immunology
MH  - beta 2-Glycoprotein I
PMC - PMC1905440
EDAT- 1999/12/14 00:00
MHDA- 1999/12/14 00:01
PMCR- 2000/12/01
CRDT- 1999/12/14 00:00
PHST- 1999/12/14 00:00 [pubmed]
PHST- 1999/12/14 00:01 [medline]
PHST- 1999/12/14 00:00 [entrez]
PHST- 2000/12/01 00:00 [pmc-release]
AID - cei1064 [pii]
AID - 10.1046/j.1365-2249.1999.01064.x [doi]
PST - ppublish
SO  - Clin Exp Immunol. 1999 Dec;118(3):480-6. doi: 10.1046/j.1365-2249.1999.01064.x.