申请人:Abbott Laboratories
公开号:US05399501A1
公开(公告)日:1995-03-21
A specific binding member such as an antigen or antibody is immobilized by covalent attachment to a solid phase such as a latex microparticle. The solid phase is reacted with a heterobifunctional or homobifunctional coupling agent to form a complex that is then reacted with a dithiol compound to form a thiolated solid phase. A specific binding member is reacted with the coupling agent to form a complex which is then reacted with the thiolated solid phase to link the specific binding member to the solid phase through thioethers. Alternatively, the dithiol compound may be reacted with the specific binding member/coupling agent complex to form a thiolated complex that is reacted with the solid phase/coupling agent complex. The coupling agent may contain a spacer. In another embodiment, the solid phase is reacted with a disulfide compound to form a complex and the complex is reacted with a reductant to form a thiolated solid phase which is reacted with a specific binding member/coupling agent complex. The immobilized specific binding member can be used in diagnostic binding assays, and it has increased sensitivity, specificity and stability, while requiring the use of less specific binding member.
一种特定的结合成员,如抗原或抗体,通过共价结合到固相上,如乳胶微粒。固相与杂二功能或同二功能偶联剂反应形成复合物,然后与二硫化合物反应形成硫化固相。特定的结合成员与偶联剂反应形成复合物,然后与硫化固相反应,通过硫醚键将特定的结合成员与固相链接。或者,二硫化合物可以与特定的结合成员/偶联剂复合物反应形成硫化复合物,然后与固相/偶联剂复合物反应。偶联剂可以包含一个间隔物。在另一种实施方式中,固相与二硫化合物反应形成复合物,然后与还原剂反应形成硫化固相,然后与特定的结合成员/偶联剂复合物反应。固定的特定结合成员可以用于诊断结合分析中,具有增加的灵敏度,特异性和稳定性,同时需要使用更少的特定结合成员。