Peek Laura J, Brey Robert N, Middaugh C Russell
Department of Pharmaceutical Chemistry, University of Kansas, 2030 Becker Drive, Lawrence, Kansas 66047, USA.
J Pharm Sci. 2007 Jan;96(1):44-60. doi: 10.1002/jps.20675.
A systematic, three-step approach was employed to develop a stable, optimized formulation of ricin toxin A-chain V76M/Y80A (rRTA) for use as a vaccine against ricin toxicity. The method first uses spectroscopic techniques to evaluate the stability of rRTA as a function of temperature and pH. To synthesize the data, empirical phase diagrams are generated to display the conditions under which the protein maintains particular conformational states. Following identification of optimal pH conditions, light scattering and fluorescence assays are employed to screen a wide variety of compounds for their abilities to stabilize rRTA. Once stabilizers were identified, the ability of rRTA to adsorb to aluminum salt adjuvants was evaluated. Desorption of the protein from the adjuvant was also analyzed. Using this approach, the optimal formulation conditions for rRTA were determined to be pH 6.0 utilizing glycerol as a stabilizer and Alhydrogel as an adjuvant. Such an approach has the potential to significantly reduce the time it takes to get vaccines into clinical testing.
采用了一种系统的三步法来开发一种稳定、优化的蓖麻毒素A链V76M/Y80A（rRTA）制剂，用作抗蓖麻毒素毒性的疫苗。该方法首先使用光谱技术评估rRTA在温度和pH值变化时的稳定性。为了综合数据，生成经验相图以展示蛋白质维持特定构象状态的条件。在确定最佳pH条件后，使用光散射和荧光测定法筛选各种化合物稳定rRTA的能力。一旦确定了稳定剂，就评估rRTA吸附到铝盐佐剂上的能力。还分析了蛋白质从佐剂上的解吸情况。使用这种方法，确定rRTA的最佳制剂条件为pH 6.0，使用甘油作为稳定剂，氢氧化铝凝胶作为佐剂。这种方法有可能显著减少疫苗进入临床试验所需的时间。
