Screening for stability and compatibility conditions of recombinant human epidermal growth factor for parenteral formulation: effect of pH, buffers, and excipients.
Screening for stability and compatibility conditions of recombinant human epidermal growth factor for parenteral formulation: effect of pH, buffers, and excipients.
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DOI：
10.1016/j.ijpharm.2013.04.054
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发表时间：
2013-08
影响因子：
5.8
通讯作者：
Hector Santana;Yaima González;P. T. Campana;Jesus Noda;Odalys Amarantes;R. Itri;A. Beldarraín;R. Páez
中科院分区：
文献类型：
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作者：
Hector Santana;Yaima González;P. T. Campana;Jesus Noda;Odalys Amarantes;R. Itri;A. Beldarraín;R. Páez
通过研究生物制药的稳定性和相容性与溶液组成的关系，可以开发出成功的胃肠外制剂。在这里，我们评估的pH值，缓冲液，离子强度，蛋白质浓度和存在的赋形剂对重组人表皮生长因子（rhEGF）的稳定性的影响。采用反相高效液相色谱法（RP-HPLC）、分子排阻色谱法（SEC-HPLC）、酶联免疫吸附试验（ELISA）、远紫外圆二色谱（CD）和光散射等方法评价其稳定性。不同方法的结果显示，在磷酸盐、Tris和组氨酸缓冲液中，在pH值接近7.0时获得了总体最大稳定性。CD结果表明，该蛋白在广泛的pH范围内是稳定的。SEC-HPLC和光散射结果表明，rhEGF在pH 6.0 - 8.0范围内聚集最小。离子强度和rhEGF浓度对反应速率常数均无显著影响。大多数rhEGF-赋形剂不稳定性发生在该蛋白质和还原糖之间。聚合物如聚乙二醇（PEG）和聚山梨醇酯增加蛋氨酸氧化。氧化和脱酰胺是rhEGF最常见的降解途径。本研究确定了开发成功的rhEGF胃肠外制剂所需考虑的关键解决方案因素。
A successful parenteral formulation can be developed by studying stability and compatibility of biopharmaceuticals as a function of solution composition. Here, we evaluate the influence of pH, buffers, ionic strength, protein concentration and presence of excipients on recombinant human epidermal growth factor (rhEGF) stability. The stability was accessed by reversed-phase high performance liquid chromatography (RP-HPLC), size exclusion chromatography (SEC-HPLC), enzyme-linked immunosorbent assay (ELISA), Far-UV circular dichroism (CD) and light scattering. The overall maximal stability was obtained in pH near to 7.0 in phosphate, Tris and histidine buffers as the results of the different methods revealed. The CD results revealed that this protein is stable in an extensive pH range. Aggregation of rhEGF was minimized at pH values ranged from 6.0 to 8.0 as indicated the SEC-HPLC and light scattering results. Nor the ionic strength neither the rhEGF concentration had significant effect on the reaction rate constants. Most rhEGF–excipient instability occurs among this protein and reducing sugars. Polymers like poly(ethylene glycol) (PEG) and polysorbates increased methionine oxidation. The rhEGF oxidation and deamidation were the most common degradation pathways. This research identified critical solution factors to be considered for the development of a successful rhEGF parenteral formulation.