Weisbjerg Paul Luigi Gargani, Caspersen Mikael Bjerg, Cook Ken, Van De Weert Marco
Novozymes Biopharma, Bagsvaerd, 2880, Denmark.
J Pharm Sci. 2015 Feb;104(2):548-56. doi: 10.1002/jps.24275. Epub 2014 Dec 1.
Aggregation levels of therapeutic proteins may be difficult to determine in mixtures containing other proteinaceous excipients. We performed a feasibility study of using serial coupling of an anion exchange and size exclusion column to determine the aggregation levels of four different model monoclonal antibodies (mAb) mixed with the model proteinaceous excipient recombinant human serum albumin (rHSA). For three of the four mAbs suitable elution conditions could be established. From the limitations imposed by the pI of the rHSA, the pI of the mAb and the nature of the columns used, it was possible to propose a set of general conditions that allows quantification of the aggregation level of a therapeutic protein in the presence of a proteinaceous excipient: The excipient protein and protein of interest should differ in pI by a minimum of 0.5 units, and the pI of the protein of interest should not be higher than ca. 8.5.
在含有其他蛋白质辅料的混合物中，治疗性蛋白质的聚集水平可能难以确定。我们进行了一项可行性研究，采用阴离子交换柱和尺寸排阻柱串联的方法来测定四种不同的模型单克隆抗体（mAb）与模型蛋白质辅料重组人血清白蛋白（rHSA）混合后的聚集水平。对于四种mAb中的三种，可以建立合适的洗脱条件。从rHSA的pI、mAb的pI以及所用柱子的性质所带来的限制来看，有可能提出一组通用条件，用于在存在蛋白质辅料的情况下定量治疗性蛋白质的聚集水平：辅料蛋白质和目标蛋白质的pI差异应至少为0.5个单位，且目标蛋白质的pI不应高于约8.5。
