Christie Merry, Peritt David, Torres Raul M, Randolph Theodore W, Carpenter John F
Department of Pharmaceutical Sciences, University of Colorado Denver, Anschutz Medical Campus, Aurora, Colorado 80045.
Global Biologics Research and Development, Hospira, Lake Forest, Illinois 60045.
J Pharm Sci. 2015 Dec;104(12):4041-4055. doi: 10.1002/jps.24639. Epub 2015 Sep 16.
Human serum albumin (HSA) is an excipient present in formulations of several recombinant protein products that are approved for clinical use. We investigated the relative contributions of HSA and HSA particles to the generation of antibody responses against recombinant human erythropoietin (rhEPO) and the excipient HSA itself. Protein samples were characterized before injection for quantities of monomeric proteins, soluble protein aggregates, and nano- and micron-sized particles. rhEPO, containing various concentrations of HSA particles, were injected three times a week for 8 weeks into mice. Hematocrits and the production of anti-rhEPO and anti-HSA antibodies were determined at various time points. Levels of antibodies against rhEPO in mice injected with HSA-containing rhEPO were higher than those in mice treated with HSA-free rhEPO. Mice injected with formulations that contained particles of HSA produced strong anti-HSA antibody responses; whereas these responses were greatly reduced when particle-free formulations were administered. In contrast, anti-rhEPO antibody responses were not affected by the presence of particles.
人血清白蛋白（HSA）是几种已获临床使用批准的重组蛋白产品制剂中的一种辅料。我们研究了HSA及其颗粒对针对重组人促红细胞生成素（rhEPO）和辅料HSA自身产生抗体反应的相对贡献。在注射前对蛋白质样品进行表征，以确定单体蛋白、可溶性蛋白聚集体以及纳米和微米级颗粒的数量。将含有不同浓度HSA颗粒的rhEPO每周注射小鼠三次，持续8周。在不同时间点测定血细胞比容以及抗rhEPO和抗HSA抗体的产生情况。注射含HSA的rhEPO的小鼠体内抗rhEPO抗体水平高于注射不含HSA的rhEPO的小鼠。注射含有HSA颗粒制剂的小鼠产生了强烈的抗HSA抗体反应；而当给予无颗粒制剂时，这些反应大大降低。相比之下，抗rhEPO抗体反应不受颗粒存在的影响。
