Capelle Martinus A H, Gurny Robert, Arvinte Tudor
Department of Pharmaceutics and Biopharmaceutics, School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Quai E-Ansermet 30, 1211, Geneva 4, Switzerland.
Pharm Res. 2009 Jan;26(1):118-28. doi: 10.1007/s11095-008-9662-8. Epub 2008 Jul 4.
The feasibility of using high throughput spectroscopy for characterization and selection of physically stable protein formulations was studied.
A hundred aqueous formulations of salmon calcitonin (sCT) were prepared using 20 buffer compositions. The solutions had pH values between 2.5 and 10.5. The stability of the sCT formulations was analyzed over 1 week by the following assays: (1) protein concentration, (2) volume control by measuring pathlength, (3) turbidity (absorbance at 350 nm), (4) intrinsic tyrosine fluorescence, (5) 1-anilino-naphthalene-8-sulfonate (ANS) fluorescence, (6) Nile Red fluorescence. Addition of the dyes (Nile Red and ANS) was used to study protein conformational changes.
After 1 day, 27 out of the 100 formulations of salmon calcitonin were stable. After 7 days, 12 stable sCT formulations remained. The best salmon calcitonin formulation was in 10 mM sodium acetate buffer with pH values between 3.5 and 5.5.
The findings are in accordance with the sCT formulations that were patented and used commercially. This can be considered as a proof of concept for the high throughput protein formulation platform.
研究使用高通量光谱法对物理稳定的蛋白质制剂进行表征和筛选的可行性。
使用20种缓冲液组合物制备了100种鲑鱼降钙素（sCT）水性制剂。溶液的pH值在2.5至10.5之间。通过以下测定法对sCT制剂的稳定性进行了为期1周的分析：（1）蛋白质浓度，（2）通过测量光程进行体积控制，（3）浊度（350nm处的吸光度），（4）内在酪氨酸荧光，（5）1-苯胺基萘-8-磺酸盐（ANS）荧光，（6）尼罗红荧光。添加染料（尼罗红和ANS）用于研究蛋白质构象变化。
1天后，100种鲑鱼降钙素制剂中有27种稳定。7天后，仍有12种稳定的sCT制剂。最佳的鲑鱼降钙素制剂是在pH值为3.5至5.5的10mM醋酸钠缓冲液中。
这些发现与已获专利并商业使用的sCT制剂一致。这可被视为高通量蛋白质制剂平台的概念验证。
