Immunogenicity of protein aggregates of a monoclonal antibody generated by forced shaking stress with siliconized and nonsiliconized syringes in BALB/c mice
Immunogenicity of protein aggregates of a monoclonal antibody generated by forced shaking stress with siliconized and nonsiliconized syringes in BALB/c mice
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DOI：
10.1111/jphp.12765
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发表时间：
2017-10-01
影响因子：
3.3
通讯作者：
Kagawa, Yoshiyuki
中科院分区：
文献类型：
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作者：
Uchino, Tomonobu;Miyazaki, Yasunori;Kagawa, Yoshiyuki
目的在本研究中，我们旨在研究在硅化和非硅化注射器强制振动应力下产生的单克隆抗体蛋白聚集体在小鼠模型中的免疫原性。方法用振荡和顶空空气分别在硅化和非硅化注射器中填充样品。表征研究采用高效粒径排除色谱、纳米颗粒跟踪分析、流式细胞术、微流成像和共振质量测量。将样品（10或100g）皮下注射BALB/c小鼠21d，监测抗药物抗体（ADA）浓度。在用硅化注射器[SO(+)]摇动的样品中，通过与硅油滴的相互作用，形成了大量亚微米和不可见的蛋白质聚集体。单抗溶液和摇匀样品的蛋白质聚集特征不同，这强烈表明硅油加速了蛋白质聚集。低剂量给药时，所有样品中的ADA浓度随着重复注射而增加，SO（+）诱导的免疫原性最高。然而，当给予高剂量时，ADA浓度在长时间重复给予耐受性后下降。结论单抗蛋白聚集可诱导小鼠免疫原性，而SO（+）诱导的免疫原性高于非硅化注射器振荡样品。
ObjectiveIn this study, we aimed to investigate the immunogenicity of protein aggregates of monoclonal antibodies (mAbs), generated by forced shaking stress with siliconized and nonsiliconized syringes in a mouse model.MethodsSamples were filled in siliconized and nonsiliconized syringes with shaking and headspace air. Characterization studies were performed using high-performance size-exclusion chromatography, nanoparticle tracking analysis, flow cytometry, micro-flow imaging and resonant mass measurement. The samples (10 or 100g) were subcutaneously injected into BALB/c mice for 21days, and the anti-drug antibody (ADA) concentrations were monitored.Key findingsIn samples shaken with siliconized syringes [SO (+)], large amounts of submicron and subvisible protein aggregates were formed by interactions with silicone oil droplets. The characteristics of protein aggregates differed between the mAb solution and shaken samples, which strongly indicates that silicone oil accelerates protein aggregation. When administered at low doses, the ADA concentration in all samples increased with repeated injections, and SO (+) induced the highest immunogenicity. However, when administered at high doses, ADA concentration decreased following prolonged repeated administration for tolerance.ConclusionsThese results indicated that mAb protein aggregation induced immunogenicity in mice, and SO (+) induced higher immunogenicity than samples shaken with nonsiliconized syringe.