Jørstad Øystein Kalsnes, Foss Stian, Gjølberg Torleif Tollefsrud, Mester Simone, Nyquist-Andersen Mari, Sivertsen Magne Sand, Fossum Dag, Gleditsch Espen, Moe Morten Carstens, Andersen Jan Terje
Department of Ophthalmology, Oslo University Hospital, Oslo, Norway.
Department of Pharmacology, Oslo University Hospital and University of Oslo, Oslo, Norway.
Int J Retina Vitreous. 2023 Nov 7;9(1):65. doi: 10.1186/s40942-023-00507-3.
Intravitreal injection (IVI) of antibody biologics is a key treatment approach in ophthalmology. Pharmaceutical compounding and storage of prefilled syringes for IVI must take place without impairing the structure and function of the biologics. This study investigated the effect of withdrawing and storing the therapeutic antibody faricimab (Vabysmo, Roche, Basel, Switzerland) in the Zero Residual silicone oil-free, 0.2-mL syringe (SJJ Solutions, The Hague, the Netherlands).
To assess the effect of syringe withdrawal on faricimab, we compared samples from syringes prepared at day 0 with samples taken directly from faricimab vials. To assess the effect of syringe storage on faricimab, we kept prefilled syringes in the dark at 4 C for 7, 14, or 37 days and compared samples from these syringes with day 0. We measured protein concentration (with spectrophotometry), stability and integrity (with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), size-exclusion chromatography (SEC), and melting temperature (Tm)), as well as binding of faricimab to its cognate antigens: vascular endothelial growth factor A (VEGF-A) and angiopoietin-2 (Ang-2) (with enzyme-linked immunosorbent assay (ELISA)).
Faricimab migrated in line with its expected molecular mass under both reducing and non-reducing conditions for all time points when analyzed with SDS-PAGE, without any sign of degradation products or aggregation. The SEC elution profiles were identical for all time points. There were slight variations in Tm for different time points compared to day 0 but without consistent relationship with storage time. ELISA did not detect differences in VEGF-A or Ang-2 binding between time points, and faricimab did not bind the neonatal Fc receptor.
Withdrawal and storage of faricimab in syringes for up to day 37 did not impair the structure and bi-specific binding properties of the therapeutic antibody.
玻璃体内注射抗体生物制剂是眼科的一种关键治疗方法。用于玻璃体内注射的预填充注射器的药物配制和储存必须在不损害生物制剂结构和功能的情况下进行。本研究调查了在零残留无硅油的0.2毫升注射器（荷兰海牙SJJ Solutions公司）中抽取和储存治疗性抗体法西单抗（Vabysmo，罗氏公司，瑞士巴塞尔）的效果。
为评估注射器抽取对法西单抗的影响，我们将第0天制备的注射器中的样品与直接从法西单抗小瓶中取出的样品进行比较。为评估注射器储存对法西单抗的影响，我们将预填充注射器在4℃黑暗中保存7、14或37天，并将这些注射器中的样品与第0天的样品进行比较。我们测量了蛋白质浓度（用分光光度法）、稳定性和完整性（用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、尺寸排阻色谱（SEC）和熔解温度（Tm）），以及法西单抗与其同源抗原血管内皮生长因子A（VEGF-A）和血管生成素-2（Ang-2）的结合（用酶联免疫吸附测定（ELISA））。
在用SDS-PAGE分析时，在所有时间点，法西单抗在还原和非还原条件下的迁移均与其预期分子量一致，没有任何降解产物或聚集的迹象。所有时间点的SEC洗脱曲线均相同。与第0天相比，不同时间点的Tm有轻微变化，但与储存时间没有一致的关系。ELISA未检测到各时间点之间VEGF-A或Ang-2结合的差异，且法西单抗不与新生儿Fc受体结合。
法西单抗在注射器中抽取和储存长达37天不会损害治疗性抗体的结构和双特异性结合特性。
