pH值对重组人源抗狂犬病病毒单克隆抗体稳定性的影响

目的研究pH值对重组人源抗狂犬病病毒单克隆抗体NM57稳定性的影响。方法将NM57原液的pH值分别调节为4.0、5.0、6.0、7.0和8.0,分别置于4和40℃保存,于0、2、4、6周取样,观察其外观的变化,并检测其SDS-PAGE及SEC-HPLC纯度。中试规模制备NM57制剂(200IU/ml,pH6.0),4℃放置3、6、9、12、18和24个月,取样观察其外观的变化,SEC-HPLC检测样品纯度,快速荧光灶抑制试验(RFFIT)检测样品中和活性。结果在观察期内,所有样品的外观均保持澄清透明。不同pH值的NM57原液4℃放置6周,纯度无明显变化;40℃放置的不同pH值的NM57原液样品纯度变化有明显差异,NM57(5.0)和NM57(6.0)样品相对稳定。中试规模制备的6批NM57制剂4℃放置24个月,其纯度及中和活性均保持稳定。结论NM57原液在甘氨酸-组氨酸缓冲系统中,pH6.0条件下比较稳定,NM57(pH6.0)制剂长期稳定性良好。

Effect of pH Value on Stability of Recombinant Human Anti-Rabies Monoclonal Antibody

Objective To study the effect of pH value on stability of recombinant human anti-rabies monoclonal antibody (McAb)NM57. Methods The pH values of bulk of McAb NM57 was adjusted to 4.0, 5.0, 6.0, 7.0 and 8.0, and stored at 4 and 40℃ separately. Samples were taken at weeks 0, 2, 4 and 6 after storage, observed for appearance and determined for purities by SDS-PAGE and SEC-HPLC. McAb NM57 (200 IU / ml, pH 6.0)was prepared on a pilot production scale and stored at 4℃ for 3, 6, 9, 12, 18 and 24 months, from which samples were taken and observed for appearance, then determined for purity by SEC-HPLC and for neutralizing activity by RFFIT. Results All the samples were clear and transparent during the whole observation period. The purities of bulk of McAb NM57 at various pH values showed no significant change after storage at 4℃ for 6 weeks, while showed significant change after storage at 40℃. The purities of bulk of McAb NM57 at pH values of 5. 0 and 6. 0 were relatively stable. Both the purities and neutralizing activities of 6 batches of McAb NM57 prepared on a pilot production scale were stable after storage at 4℃ for 24 months. Conclusion The bulk McAb NM57 was stable at pH 6.0 in glycine-histidine buffer system, and the final product (pH 6.0)showed good long-term stability.