Nygren H, Hansson H A, Lange S
Med Biol. 1979 Jun;57(3):187-91.
Horseradish peroxidase was conjugated to immunoglobulin G via glutaraldehyde by a two-step procedure using an increasing excess of peroxidase in the second step reaction. The yield of conjugated monomeric IgG and the amount of free IgG were analyzed by SDS-polyacrylamide electrophoresis and gel-filtration. The antigen binding capacity of the enzyme-antibody conjugates was evaluated by radial immunodiffusion. Conjugation of peroxidase to IgG with a 1:20 molar:molar excess of glutaraldehyde-activated peroxidase resulted in a high yield of conjugated IgG without any detectable amounts of polymers of IgG or residual free IgG. The antigen binding capacity of the conjugate varied between different antigen-antibody systems, but in general it was not significantly different from that of native IgG. The enzyme activity was reduced to 70% of the activity of native peroxidase.
辣根过氧化物酶通过戊二醛与免疫球蛋白G偶联，采用两步法，在第二步反应中使用逐渐增加的过量过氧化物酶。通过SDS-聚丙烯酰胺凝胶电泳和凝胶过滤分析偶联的单体IgG的产量和游离IgG的量。通过放射免疫扩散评估酶-抗体偶联物的抗原结合能力。用1:20摩尔比过量的戊二醛活化过氧化物酶将过氧化物酶与IgG偶联，得到高产率的偶联IgG，未检测到IgG聚合物或残留游离IgG。偶联物的抗原结合能力在不同的抗原-抗体系统之间有所不同，但总体上与天然IgG没有显著差异。酶活性降低到天然过氧化物酶活性的70%。
