Nagasawa J, Suehiro T, Yamauchi A
J Appl Biochem. 1985 Dec;7(6):430-7.
The preparation of adsorbents for DNA antibodies is described. The degree of immobilization of native DNA on Sepharoses activated with epichlorohydrin or bisoxirane was investigated as a function of pH, temperature, time, concentration of DNA, and oxirane content in the supports. The maximum amount of DNA bound was obtained after 8 h at 40-50 degrees C at pH 11-11.5. The amount bound was increased by raising either the concentration of DNA or the oxirane content of the supports, and could reach 300 mg/g dry support. The immobilized DNA was applied to the adsorption of DNA antibodies using either commercial human serum with anti-native DNA activity or the sera of patients with systemic lupus erythematosus. The amount of antibody adsorbed depended on the amount of DNA. The thermal stability of the immobilized DNA was also examined. After heating at 80 degrees C, the leakage of DNA was slight and the adsorption of antibodies was not affected.
本文描述了用于DNA抗体的吸附剂的制备方法。研究了天然DNA在环氧氯丙烷或双环氧乙烷活化的琼脂糖上的固定化程度与pH值、温度、时间、DNA浓度以及载体中环氧乙烷含量的关系。在40 - 50℃、pH值为11 - 11.5的条件下反应8小时后,DNA的结合量达到最大值。提高DNA浓度或载体中环氧乙烷含量均可增加结合量,最高可达300 mg/g干载体。使用具有抗天然DNA活性的商业人血清或系统性红斑狼疮患者的血清,将固定化的DNA用于吸附DNA抗体。吸附的抗体量取决于DNA的量。还检测了固定化DNA的热稳定性。在80℃加热后,DNA的泄漏量很少,且抗体的吸附不受影响。