Sesardic D, Prior C, Daas A, Buchheit K H
National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK.
Pharmeuropa Bio. 2003 Jul;2003(1):5-21.
A stable liquid candidate Biological Reference Preparation (BRP) for diphtheria toxin was prepared in peptone buffer (nominal content of diphtheria toxin: 1 Lf/ml, 0.4 micro g/ml), filled in ampoules (filling volume: 1 ml) and characterised in a collaborative study. The toxin is to be used in the test "Absence of toxin and irreversibility of toxoid" as described in the current European Pharmacopoeia (Ph. Eur.) monograph Diphtheria Vaccine (Adsorbed) (2002:0443). Eleven laboratories assessed the specific activity of the preparation by in vivo and in vitro assays. The material is assumed to have satisfactory stability with a calculated predicted loss of activity of <1% per year at 4-8 degrees C. From the collaborative study, the specific activity was calculated as 77.6 (45-113) LD( 50)/ml (lethal challenge) and >75 000 Lr/Lf (intradermal challenge). The candidate BRP was successfully used in nine laboratories and confirmed suitable for use in the Vero cell test for "Absence of toxin and irreversibility of toxoid" as described in the Ph. Eur. monograph 2002:0443; i.e., concentrations of 5 x 10( -5) Lf/ml and below caused cytotoxic effects in the Vero cell test. Due to its liquid nature, the stability of the material will be monitored at regular intervals and preparation of a stable freeze-dried formulation will be considered for long-term use. Additional studies will be performed to confirm suitability of this BRP for other applications. The candidate BRP was adopted as the Ph. Eur. reference material for Diphtheria Toxin Batch 1 by the Ph. Eur. Commission at its session in March 2003.
制备了一种用于白喉毒素的稳定液体候选生物参考制剂（BRP），该制剂采用蛋白胨缓冲液配制（白喉毒素标称含量：1Lf/ml，0.4μg/ml），灌装于安瓿中（灌装体积：1ml），并在一项协作研究中进行了特性鉴定。该毒素将用于现行欧洲药典（Ph. Eur.）专论《吸附白喉疫苗》（2002:0443）中所述的“毒素不存在及类毒素不可逆性”试验。11个实验室通过体内和体外试验评估了该制剂的比活性。假定该材料具有令人满意的稳定性，在4-8℃下计算得出的预测活性损失<1%/年。根据协作研究，比活性计算为77.6（45-113）LD(50)/ml（致死攻击）和>75000Lr/Lf（皮内攻击）。候选BRP在9个实验室中成功使用，并确认适用于Ph. Eur.专论2002:0443中所述的“毒素不存在及类毒素不可逆性”的Vero细胞试验；即，5×10(-5)Lf/ml及以下的浓度在Vero细胞试验中引起细胞毒性作用。由于其液体性质，将定期监测该材料的稳定性，并考虑制备稳定的冻干制剂以供长期使用。将进行额外研究以确认该BRP适用于其他用途。2003年3月，Ph. Eur.委员会会议将候选BRP采纳为Ph. Eur.白喉毒素第1批参考物质。
