Voronov A V, Gudima S O, Mierinia A A, Vengerov Iu Iu, Votrin I I
Biull Eksp Biol Med. 1988 Aug;106(8):156-9.
Four different chromatographic methods of IgG isolation from rabbit antisera to placental alkaline phosphatase (HPAP) have been compared. The antibodies were obtained by ion-exchange chromatography and affinity chromatography on protein-A-sepharose, on the sepharose with immobilized antigen. IgG samples were characterized by the content of specific antibodies to HPAP and checked in enzyme immunoassay (EIA). IgG purified on immobilized antigen were found to be the optimal both from the point of view of the specific antibodies content and EIA sensitivity, but satisfactory results could be also obtained with ion-exchange and protein-A-chromatography purified IgG. The last two isolation methods are simpler and provide 3-10 ng/ml sensitivity of HPAP detection, which is lower, as compared with the test employing affinity antibodies (1 ng/ml), but allows the detection of HPAP in serum samples.
比较了从兔抗胎盘碱性磷酸酶（HPAP）抗血清中分离IgG的四种不同色谱方法。通过离子交换色谱法以及在蛋白A-琼脂糖、固定化抗原琼脂糖上进行亲和色谱法获得抗体。IgG样品通过针对HPAP的特异性抗体含量进行表征，并在酶免疫测定（EIA）中进行检测。从特异性抗体含量和EIA灵敏度的角度来看，在固定化抗原上纯化的IgG被发现是最优的，但使用离子交换和蛋白A色谱法纯化的IgG也能获得令人满意的结果。最后两种分离方法更简单，提供了3 - 10 ng/ml的HPAP检测灵敏度，与使用亲和抗体的检测方法（1 ng/ml）相比更低，但能够检测血清样品中的HPAP。
