Abstract:
This invention presents novel assay methods employing a capture reagent, involving a first binding member typically conjugated to a polymeric anion substance, and a solid phase material containing a reaction site comprising a polymeric cation substance having a nitrogen content of at least about two percent. In one embodiment a test sample suspected of containing the analyte of interest may be contacted with the capture reagent to form a charged capture reagent/analyte complex. The complex is then contacted to the oppositely charged solid phase to attract, attach, and immobilize the capture reagent/analyte complex. The use of a polycationic substance, having a nitrogen content of at least about two percent (excluding the counter ions), to prepare a suitably charged solid phase provides a solid phase that can be subjected to a greater extent of manipulation, such as multiple washes, without losing the capability to attract and retain the capture reagent.
Abstract:
The present invention includes novel assays and reagents using a capture reagent, involving a specific binding member attached to a polymeric anionic substance, and a solid phase material containing a capture or reaction zone including a polymeric cationic substance. A test sample suspected of containing the analyte of interest may be contacted with the capture reagent to form a charged capture reagent/analyte complex. The complex is then contacted to the oppositely charged solid phase to attract, attach, and immobilize the capture reagent/analyte complex. With an appropriate indicator reagent, sandwich, competitive and indirect assays can be performed. Preferably, the indicator reagent includes a nonspecific binding blocker to reduce the nonspecific binding between the indicator reagent and the solid phase, without interfering with the reaction between the capture reagent and the solid phase. Suitable nonspecific binding blockers include unbound or free polyanionic materials such as dextran sulfate, heparin, carboxymethyl dextran, carboxymethyl cellulose, pentosan polysulfate, inositol hexasulfate or beta -cyclodextrin sulfate.
Abstract:
Nouveaux procédés d'analyse et réactif faisant appel à un réactif de capture, mettant en jeu un membre de fixation spécifique associé à une substance anionique polymère, et un matériau en phase solide contenant une zone de capture ou de réaction comportant une substance cationique polymère. Un échantillon à analyser dans lequel on soupçonne la présence de l'analyte recherché peut être mis en contact avec le réactif de capture pour former un complexe chargé contenant le réactif de capture et l'analyte. Le complexe est ensuite mis en contact avec la phase solide de charge opposée pour attirer, fixer et immobiliser le complexe réactif de capture/analyte. Avec un réactif indicateur approprié, on peut réaliser des analyses en sandwich, compétitives et indirectes. De préférence, le réactif indicateur comprend un inhibiteur de fixation non spécifique qui permet de réduire la fixation non spécifique entre les réactifs indicateurs et la phase solide, sans gêner la réaction entre le réactif de capture et la phase solide. Les inhibiteurs de fixation non spécifiques appropriés comprennent des matériaux polyanioniques non liés ou libres tels que le sulfate de dextrane, l'héparine, le dextrane de carboxyméthyle, la cellulose de carboxyméthyle, le polysulfate de pentosane, l'hexasulfate d'inositol ou le sulfate de beta-cyclodextrine.
Abstract:
Nouveaux procédés d'analyse faisant appel à un réactif de capture, mettant en jeu un premier membre de fixation normalement associé à une substance anionique polymérique, un matériau en phase solide contenant un lieu de réaction constitué d'une substance cationique polymérique d'une teneur en azote d'au moins environ 2 %. Dans un mode de réalisation, un échantillon à analyser dans lequel on soupçonne la présence de l'analyte recherché peut être mis en contact avec le réactif de capture pour former un complexe chargé contenant le réactif de capture et l'analyte. Le complexe est ensuite mis en contact avec la phase solide de charge opposée pour attirer, fixer et immobiliser le complexe réactif de capture/analyte. L'utilisation d'une substance polycationique, d'une teneur en hydrogène d'au moins environ 2 % (à l'exclusion des contre-ions), dans la préparation d'une phase solide correctement chargée fournit une phase solide qui peut être soumise à un plus grande nombre de manipulations, tels que les lavages multiples, sans perdre la capacité d'attirer et de retenir le réactif de capture.
Abstract:
This invention presents novel assay methods employing a capture reagent, involving a first binding member typically conjugated to a polymeric anion substance, and a solid phase material containing a reaction site comprising a polymeric cation substance having a nitrogen content of at least about two percent. In one embodiment a test sample suspected of containing the analyte of interest may be contacted with the capture reagent to form a charged capture reagent/analyte complex. The complex is then contacted to the oppositely charged solid phase to attract, attach, and immobilize the capture reagent/analyte complex. The use of a polycationic substance, having a nitrogen content of at least about two percent (excluding the counter ions), to prepare a suitably charged solid phase provides a solid phase that can be subjected to a greater extent of manipulation, such as multiple washes, without losing the capability to attract and retain the capture reagent.
Abstract:
The present invention includes novel assays and reagents using a capture reagent, involving a specific binding member attached to a polymeric anionic substance, and a solid phase material containing a capture or reaction zone including a polymeric cationic substance. A test sample suspected of containing the analyte of interest may be contacted with the capture reagent to form a charged capture reagent/analyte complex. The complex is then contacted to the oppositely charged solid phase to attract, attach, and immobilize the capture reagent/analyte complex. With an appropriate indicator reagent, sandwich, competitive and indirect assays can be performed. Preferably, the indicator reagent includes a nonspecific binding blocker to reduce the nonspecific binding between the indicator reagent and the solid phase, without interfering with the reaction between the capture reagent and the solid phase. Suitable nonspecific binding blockers include unbound or free polyanionic materials such as dextran sulfate, heparin, carboxymethyl dextran, carboxymethyl cellulose, pentosan polysulfate, inositol hexasulfate or beta -cyclodextrin sulfate.