Solid phase reagent and assay method for measuring antibodies specific to antiphospholipid syndrome
AssigneesYamasa Corporation, Takao Koike
InventorsEiji Matsuura, Yoshiko Igarashi, Hisato Nagae, Takao Koike
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Abstract
A solid phase reagent is provided which comprises a carrier having a surface onto which functional groups containing a negative charge or a lone pair of electrons and/or free radicals containing a negative charge or a lone pair of electrons have been introduced and having .beta.2-glycoprotein I coated on the surface. Using this reagent, antibodies specific to antiphospholipid syndrome can be specifically assayed. Moreover, antibodies specific to antiphospholipid syndrome can be assayed differentially from antibodies specific to infectious diseases by using the solid phase reagent described above or a further modified solid phase reagent.
Claims15
A solid phase reagent for detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome, which reagent does not contain a phospholipid, said reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, and which resin further has .beta.2-glycoprotein I bound to said derivatized surface.
A method for detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome, which method does not utilize a phospholipid, said method consisting essentially of the steps of:
- contacting a fluid test sample with a solid phase reagent, said reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, and which resin further has .beta.2-glycoprotein I bound to said derivatized surface, to specifically bind anticardiolipin antibodies in the fluid test sample which are specific to antiphospholipid syndrome to the .beta.2-glycoprotein I of the solid phase reagent, and
- detecting the specifically bound anticardiolipin antibodies.
A kit for use in detecting anticardiolipin antibodies in a fluid test sample which are specific to antiphospholipid syndrome, which kit does not contain a phospholipid, said kit comprising:
- a solid phase reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, and which resin further has .beta.2-glycoprotein I bound to said derivatized surface, and
- means for detecting anticardiolipin antibodies bound to the solid phase reagent which are specific to antiphospholipid syndrome, which means do not contain a phospholipid.
A solid phase reagent for detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome, which reagent does not contain a phospholipid, said reagent consisting essentially of a synthetic resin having a protein-adsorbable hyrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, said derivatized surface having a plurality of binding sites for .beta.2-glycoprotein I and wherein .beta.2-glycoprotein I is bound to only a portion of said plurality of binding sites on said derivatized surface.
A method for selectively detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome differentially from anticardiolipin antibodies which are specific to infectious diseases, which method does not utilize a phospholipid, said method consisting essentially of the steps of:
- contacting a fluid test sample with a solid phase reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, said derivatized surface having a plurality of binding sites for .beta.2-glycoprotein I, and wherein.beta.2-glycoprotein I is bound to only a portion of said plurality of binding sites on said derivatized surface,
- wherein anticardiolipin antibodies in the fluid test sample which are specific to antiphospholipid syndrome are specifically bound to the .beta.2-glycoprotein I of the solid phase reagent, whereas anticardiolipin antibodies in the fluid test sample which are specific to infectious diseases are bound to the solid phase reagent independently of .beta.2-glycoprotein I.
- and comparing the extent of binding by anticardiolipin antibodies in the fluid test sample to said derivatized surface having .beta.2-glycoprotein I bound thereto with the extent of binding by anticardiolipin antibodies in the fluid test sample to said derivatized surface having no .beta.2-glycoprotein I bound thereto, thereby to differentially determine anticardiolipin antibodies in the fluid test sample which are specific to antiphospholipid syndrome from anticardiolipin antibodies which are specific to infectious diseases.
A kit for use in differentially detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome from anticardiolipin antibodies which are specific to infectious diseases, which kit does not contain a phospholipid, said kit comprising:
- a solid phase reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, said derivatized surface having a plurality of binding sites for .beta.2-glycoprotein I. and wherein .beta.2-glycoprotein I is bound to only a portion of said plurality of binding sites on said derivatized surface, and
- means for differentially detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome from anticardiolipin antibodies which are specific to infectious diseases, which means do not contain a phospholipid.
A method for selectively detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome differentially from anticardiolipid antibodies which are specific to infectious diseases, which method does not utilize a phospholipid, said method consisting essentially of the steps of:
- contacting a fluid test sample with first and second solid phase reagents, said first solid phase reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, and which resin further has .beta.2-glycoprotein I bound to said derivatized surface, said second solid phase reagent being substantially identical to said first solid phase reagent except that it has no .beta.2-glycoprotein I bound to said derivatized surface,
- wherein anticardiolipin antibodies in the fluid test sample which are specific to antiphospholipid syndrome are specifically bound to the .beta.2-glycoprotein I of said first solid phase reagent, whereas anticardiolipin antibodies in the fluid test sample which are specific to infectious diseases are bound to said solid phase reagents independently of .beta.2-glycoprotein I.
- and comparing the extent of binding by anticardiolipin antibodies in the fluid test sample to said derivatized surface of said first solid phase reagent having .beta.2-glycoprotein I bound thereto with the extent of binding by anticardiolipin antibodies in the fluid test sample to said derivatized surface of said second solid phase reagent having no .beta.2-glycoprotein I bound thereto, thereby to differentially determine anticardiolipin antibodies in the fluid test sample which are specific to antiphospholipid syndrome from anticardiolipin antibodies which are specific to infectious diseases.
A kit for use in differentially detecting anticardiolipin antibodies specific to antiphospholipid syndrome from anticardiolipin antibodies specific to infectious diseases, which kit does not contain a phospholipid, said kit comprises:
- a first solid phase reagent consisting essentially of a synthetic resin having a protein-adsorbable hydrophobic surface which is derivatized with at least one group consisting of functional groups and free radicals, said functional groups and free radicals having a negative charge, a lone pair of electrons, or both, provided that said functional groups and free radicals are not phospholipids, and which resin further has .beta.2-glycoprotein I bound to said derivatized surface,
- a second solid phase reagent which is substantially identical to said first solid phase reagent except that it has no .beta.2-glycoprotein I bound to said derivatized surface, and
- means for differentially detecting anticardiolipin antibodies which are specific to antiphospholipid syndrome from anticardiolipin antibodies which are specific to infectious diseases, which means do not contain a phospholipid.
Dependencies are inferred from the claim text.