Seematter R, Jaquet H
Ann Immunol (Paris). 1976 Jan-Feb;127(1):3-10.
The antigen binding capacity (ABC) of mouse and rabbit antisera to horseradish peroxidase (HRP) was measured by the Farr technique, originally introduced for bovine serum albumin. Results obtained by this method are independent of other antibody interactions (precipitation, formation of immune complexes, effect on enzyme activity). Commercially available HRP has variable solubility in ammonium sulfate solutions; almost 40% was precipitated at 50% saturation. Accordingly, the fraction of HRP soluble at 60% saturation was used, and this was labelled with 125I2 using the HRP oxidizing activity. Under these test conditions, accurate and reproducible results were obtained with protein concentrations (as N/ml) of the order to 0.07 mug (antibody) and 0.01 mug (antigen) respectively. This high sensitivity is superior to that achieved by passive hemagglutination, especially when titrating antibody levels in mouse sera.
采用最初用于牛血清白蛋白的Farr技术测定了小鼠和兔抗血清与辣根过氧化物酶（HRP）的抗原结合能力（ABC）。通过该方法获得的结果与其他抗体相互作用（沉淀、免疫复合物形成、对酶活性的影响）无关。市售的HRP在硫酸铵溶液中的溶解度各不相同；在50%饱和度时，几乎40%会沉淀。因此，使用了在60%饱和度下可溶的HRP部分，并利用HRP的氧化活性用125I2对其进行标记。在这些测试条件下，蛋白质浓度（以N/ml计）分别为0.07微克（抗体）和0.01微克（抗原）时，可获得准确且可重复的结果。这种高灵敏度优于被动血凝试验所达到的灵敏度，尤其是在滴定小鼠血清中的抗体水平时。
