Derango R, Page J
Hybritech Incorporated, San Diego, CA 92121, USA.
J Immunoassay. 1996 May;17(2):145-53. doi: 10.1080/01971529608005785.
Quantitation of antibody coupled to a derivatized polystyrene bead through a bifunctional cross linker can be accomplished by a competitive enzyme-linked immunosorbent assay (ELISA) method. This sensitive method is less subject to interference than other protein assay methods such as bicinchoninic acid (BCA) or Lowry. The competitive ELISA method consists of incubating the coupled bead with a (20/80) weight ratio of goat anti mouse kappa alkaline phosphatase/goat anti mouse kappa (GAMKAP/GAMK) for 1.5 hours at 37 degrees C, washing, adding p-nitrophenyl phosphate (PNPP) substrate, and reading the absorbance at 405/450 nm. A standard curve is established with radiolabeled antibody beads for microgram quantitation.
通过双功能交联剂与衍生化聚苯乙烯珠偶联的抗体的定量可通过竞争性酶联免疫吸附测定（ELISA）方法来完成。这种灵敏的方法比其他蛋白质测定方法（如二辛可宁酸（BCA）或洛里法）受干扰的影响更小。竞争性ELISA方法包括将偶联的珠子与山羊抗小鼠κ碱性磷酸酶/山羊抗小鼠κ（GAMKAP/GAMK）以（20/80）重量比在37℃下孵育1.5小时，洗涤，加入对硝基苯磷酸酯（PNPP）底物，并读取405/450nm处的吸光度。用放射性标记的抗体珠子建立标准曲线用于微克定量。
