Sato H, Kidaka T, Hori M
Medical Supply and Equipment Department, Takeda Chemical Industries, Ltd., Osaka, Japan.
Appl Biochem Biotechnol. 1987 Aug;15(2):145-58. doi: 10.1007/BF02801315.
In developing therapeutic immunoadsorbents (IAs), antibodies (IgG molecules) covalently immobilized on porous carriers, a leak of IgG was determined both in the storage test with buffers at 25 and 4 degrees C and in contact with plasma at room temperature (RT). The amount of antibody released from therapeutic IAs must be minimized to avoid side effects during treatment. The amount of IgG released was a. Dependent on the amount of IgG immobilized b. Much greater with CNBr-activated Sepharose 4B, or Formyl-Cellulofine as a support material than with aminopropyl CPG (controlled pore glass) c. Found to yield again during another storage in buffers after the IAs were washed and their buffers replaced with fresh ones and d. Decreased after the IAs were treated with glutaraldehyde (GA) solutions. Whereas treating the IAs with GA solutions significantly reduced the amount of IgG released, it caused some deterioration of the adsorption characteristics of the IAs. An irradiation dose of 2.5 Mrad as a crosslinking procedure also reduced the amount of IgG released; its effect was comparable to that of 0.025% GA, the lowest concentration used.
在开发治疗性免疫吸附剂(IA)时,即抗体(IgG分子)共价固定在多孔载体上,在25℃和4℃缓冲液储存试验以及室温(RT)下与血浆接触时,均测定到了IgG泄漏。治疗性IA释放的抗体量必须降至最低,以避免治疗期间出现副作用。释放的IgG量:a. 取决于固定的IgG量;b. 以溴化氰活化的琼脂糖4B或甲酰化纤维素微球作为载体材料时,比以氨丙基CPG(可控孔径玻璃)作为载体材料时释放量要大得多;c. 在IA洗涤并用新鲜缓冲液替换原有缓冲液后再次储存时,发现释放量会再次出现;d. 在IA用戊二醛(GA)溶液处理后释放量减少。虽然用GA溶液处理IA可显著降低IgG释放量,但会导致IA的吸附特性有所恶化。作为交联程序的2.5兆拉德辐照剂量也减少了IgG释放量;其效果与所用最低浓度0.025%的GA相当。