Griswold W R
J Immunol Methods. 1981;40(1):73-8. doi: 10.1016/0022-1759(81)90081-8.
Antibody bound in an antigen-antibody complex was barely detectable by the standard Farr technique using 10 microgram/ml of radiolabeled antigen. When the Farr test was repeated using larger concentrations of antigen the ABC measurements increased several-fold. In separate experiments complexes were dissociated with low pH buffer. The radiolabeled antigen was then added and the mixture returned to neutral pH before the Farr assay was performed. Pretreatment of the complexes with citrate-buffered saline pH 3.1 enhanced antibody measurements 1.5-3.6-fold. Pretreatment with glycine-buffered saline pH 2.2 enhanced complexed antibody measurements 2.1-4.8-fold. The low pH buffers did not affect ABC values on free antibody. Antibody bound in immune complexes can be detected with a modified Farr technique using a high concentration of antigen and/or by pretreatment of the complexes with low pH buffers before the addition of radiolabeled antigen.
采用标准的法尔氏技术,使用10微克/毫升的放射性标记抗原,几乎检测不到结合在抗原 - 抗体复合物中的抗体。当使用更高浓度的抗原重复进行法尔氏试验时,抗原结合抗体(ABC)测量值增加了几倍。在单独的实验中,用低pH缓冲液解离复合物。然后加入放射性标记抗原,并在进行法尔氏测定之前将混合物恢复至中性pH。用pH 3.1的柠檬酸盐缓冲盐水预处理复合物可使抗体测量值提高1.5至3.6倍。用pH 2.2的甘氨酸缓冲盐水预处理可使结合在复合物中的抗体测量值提高2.1至4.8倍。低pH缓冲液不影响游离抗体的ABC值。结合在免疫复合物中的抗体可以通过改良的法尔氏技术,使用高浓度抗原和/或在加入放射性标记抗原之前用低pH缓冲液预处理复合物来检测。