Kato K, Hamaguchi Y, Okawa S, Ishikawa E, Kobayashi K
J Biochem. 1977 Jul;82(1):261-6. doi: 10.1093/oxfordjournals.jbchem.a131678.
Rabbit antibody IgG was bound onto aminoalkylsilyl or plain glass rods by simple adsorption. For comparison, rabbit antibody IgG was also bound onto glutaraldehyde-activated aminoalkylsilyl glass rods. These antibody-glass rods were tested by the sandwich procedure using Fab' fragments of rabbit antibody conjugated with beta-D-galactosidase from Escherichia coli. The glutaraldehyde-activated aminoalkylsilyl glass showed the largest capacity to bind antigen and the plain glass showed the smallest. However, the antibody-glass rods prepared by simple adsorption were as useful for the sandwich immunoassay of macromolecular antigens as those prepared with glutaraldehyde. With all the antibody-glass rods prepared, 0.1 to 10 fmol of ornithine delta-aminotransferase from rat liver and 2,4-dinitrophenyl human IgG were measurable. More than 10 fmol of the antigens may be measurable with larger amounts of the antibody-beta-D-galactosidase complexes, although the non-specific binding of the complexes to the solid phase increases to limit the sensitivity of the immunoassay.
兔抗体IgG通过简单吸附作用结合到氨烷基硅烷化玻璃棒或普通玻璃棒上。作为对照，兔抗体IgG也结合到戊二醛活化的氨烷基硅烷化玻璃棒上。这些抗体玻璃棒通过夹心程序进行检测，该程序使用与来自大肠杆菌的β-D-半乳糖苷酶偶联的兔抗体Fab'片段。戊二醛活化的氨烷基硅烷化玻璃显示出结合抗原的最大能力，而普通玻璃显示出最小的能力。然而，通过简单吸附制备的抗体玻璃棒对于大分子抗原的夹心免疫测定与用戊二醛制备的那些一样有用。对于所制备的所有抗体玻璃棒，大鼠肝脏中的0.1至10 fmol鸟氨酸δ-氨基转移酶和2,4-二硝基苯基人IgG都是可测量的。尽管复合物与固相的非特异性结合增加会限制免疫测定的灵敏度，但使用大量的抗体-β-D-半乳糖苷酶复合物可能可测量超过10 fmol的抗原。
