Effect of glass primary container surface modification on monoclonal antibody aggregates induced by mechanical stress

doi:10.11949/0438-1157.20230201

Abstract

Abstract:

The inner surface of middle borosilicate glass tubing was modified by octadecyltrichlorosilane (OTS). The contact angles of modified surfaces which were measured by contact angle goniometer increased from 50°±1° to 90°±2°. The surface roughness which was measured by atomic force microscope increased from 0.448±0.086 to 1.282±0.117, and the result of Fourier transform infrared spectroscopy (FTIR) indicated that the OTS was successfully coated on the glass surfaces. The monoclonal antibody in glass containers were subjected to mechanical stress. Microflow imaging, size exclusion-high performance liquid chromatography (SE-HPLC) and extrinsic fluorescence dye were used to characterize the stability of monoclonal antibody. The results showed that the OTS-treated hydrophobic interface could reduce the generation of mechanical stress-induced protein aggregates. Due to the strong hydrophobic interaction between hydrophobic surface and protein, the hydrophobic surface could effectively reduce the aggregation of antibody molecules induced by mechanical stress.

Key words: surface chemistry, primary container, absorption, antibody, aggregation

摘要：

利用十八烷基三氯硅烷（octadecyltrichlorosilane，OTS）对中硼硅玻璃管制注射剂瓶内表面进行化学改性，使用接触角仪（水接触角从50°±1°增加至90°±2°）、原子力显微镜（表面粗糙度从0.448±0.086增加至1.282±0.117）、红外（出现—CH₂—和—CH₃特征峰）对表面进行验证，成功制备了具有疏水性能的玻璃表面。给玻璃瓶中的单克隆抗体施加机械应力，使用微流成像技术、尺寸排阻-高效液相色谱法（size exclusion-high performance liquid chromatography，SE-HPLC）、外源荧光对单抗制剂的稳定性进行全面表征。结果表明，OTS处理的疏水性界面可以减少机械应力诱导的蛋白质聚集体的产生。由于疏水表面与蛋白质强的疏水相互作用，OTS处理的疏水表面可有效抑制机械应力诱导的抗体分子的聚集。

关键词: 表面化学, 内包材, 吸附, 抗体, 聚集

CLC Number:

TQ 464.9

Xinyue WANG, Junjie WANG, Sixian CAO, Cui WANG, Lingkun LI, Hongyu WU, Jing HAN, Hao WU. Effect of glass primary container surface modification on monoclonal antibody aggregates induced by mechanical stress[J]. CIESC Journal, 2023, 74(6): 2580-2588.

王新悦, 王俊杰, 曹思贤, 王翠, 李灵坤, 吴宏宇, 韩静, 吴昊. 玻璃内包材界面修饰对机械应力诱导的单克隆抗体聚集体形成的影响[J]. 化工学报, 2023, 74(6): 2580-2588.

Figures/Tables 10

Fig.1 Flow chart of chemical reaction of OTS on glass surface

Fig.2 Results of water contact angle

Fig.3 FTIR spectrum of glass surfaces

Fig.4 Three-dimensional surface topography of AFM

Fig.5 Langmuir adsorption model of IgG2 on glass surfaces

Fig.6 IgG2 protein aggregates produced by mechanically stressed in untreated and OTS glass vials

Fig.7 Size distribution of subvisible particles in monoclonal antibody after mechanical stress treatment

Fig.8 Morphology of subvisible particles in monoclonal antibody solution after mechanical stress treatment in untreated glass vials and glass vials coated by 17 μmol/L OTS

Fig.9 Contour plots of IgG2 subvisible particle size and circularity

Fig.10 SE-HPLC spectrum of soluble IgG2

References 36

1	Nuévalos M, García-Ríos E, Mancebo F J, et al. Novel monoclonal antibody-based therapies: implications for the treatment and prevention of HCMV disease[J]. Trends in Microbiology, 2023, 31(5): 480-497.
2	Chen Y, Zhang G L, Yang Y W, et al. The treatment of inflammatory bowel disease with monoclonal antibodies in Asia[J]. Biomedicine & Pharmacotherapy, 2023, 157: 114081.
3	Chi E Y, Krishnan S, Randolph T W, et al. Physical stability of proteins in aqueous solution: mechanism and driving forces in nonnative protein aggregation[J]. Pharmaceutical Research, 2003, 20(9): 1325-1336.
4	Kosloski M P, Miclea R D, Balu-Iyer S V. Role of glycosylation in conformational stability, activity, macromolecular interaction and immunogenicity of recombinant human factor Ⅷ[J]. The AAPS, Journal, 2009, 11(3): 424-431.
5	Johann F, Wöll S, Winzer M, et al. Miniaturized forced degradation of therapeutic proteins and ADCs by agitation-induced aggregation using orbital shaking of microplates[J]. Journal of Pharmaceutical Sciences, 2022, 111(5): 1401-1413.
6	金鹤. 静脉输液中不溶性微粒对人体的危害及控制[J]. 上海护理, 2007, 7(5): 55-57.
	Jin H. Harm and control of insoluble particles in intravenous infusion to human body[J]. Shanghai Nursing, 2007, 7(5): 55-57.
7	Joubert M K, Hokom M, Eakin C, et al. Highly aggregated antibody therapeutics can enhance the in vitro innate and late-stage T-cell immune responses[J]. Journal of Biological Chemistry, 2012, 287(30): 25266-25279.
8	郭莎, 贾哲, 吴昊, 等. 单克隆抗体颗粒表征的现状与挑战[J]. 中国药事, 2022, 36(2)161-169.
	Guo S, Jia Z, Wu H, et al. Current status and challenges of particle characterization in monoclonal antibody formulation[J]. Chinese Pharmaceutical Affairs, 2022, 36(2)161-169.
9	Patapoff T W, Esue O. Polysorbate 20 prevents the precipitation of a monoclonal antibody during shear[J]. Pharmaceutical Development and Technology, 2009, 14(6): 659-664.
10	Bam N B, Cleland J L, Yang J, et al. Tween protects recombinant human growth hormone against agitation-induced damage via hydrophobic interactions[J]. Journal of Pharmaceutical Sciences, 1998, 87(12): 1554-1559.
11	Wang W, Wang Y J, Wang D Q. Dual effects of Tween 80 on protein stability[J]. International Journal of Pharmaceutics, 2008, 347(1/2): 31-38.
12	Perevozchikova T, Nanda H, Nesta D P, et al. Protein adsorption, desorption, and aggregation mediated by solid-liquid interfaces[J]. Journal of Pharmaceutical Sciences, 2015, 104(6): 1946-1959.
13	Kaivosoja E, Barreto G, Levón K, et al. Chemical and physical properties of regenerative medicine materials controlling stem cell fate[J]. Annals of Medicine, 2012, 44(7): 635-650.
14	Wu H, Randolph T W. Aggregation and particle formation during pumping of an antibody formulation are controlled by electrostatic interactions between pump surfaces and protein molecules[J]. Journal of Pharmaceutical Sciences, 2020, 109(4): 1473-1482.
15	Movafaghi S, Wu H, Francino Urdániz I M, et al. The effect of container surface passivation on aggregation of intravenous immunoglobulin induced by mechanical shock[J]. Biotechnology Journal, 2020, 15(9): e2000096.
16	Yoneda S, Maruno T, Mori A, et al. Influence of protein adsorption on aggregation in prefilled syringes[J]. Journal of Pharmaceutical Sciences, 2021, 110(11): 3568-3579.
17	Biddlecombe J G, Craig A V, Zhang H, et al. Determining antibody stability: creation of solid-liquid interfacial effects within a high shear environment[J]. Biotechnology Progress, 2007, 23(5): 1218-1222.
18	Oliva A, Santoveña A, Fariña J, et al. Effect of high shear rate on stability of proteins: kinetic study[J]. Journal of Pharmaceutical and Biomedical Analysis, 2003, 33(2): 145-155.
19	Sediq A S, van Duijvenvoorde R B, Jiskoot W, et al. No touching!Abrasion of adsorbed protein is the root cause of subvisible particle formation during stirring[J]. Journal of Pharmaceutical Sciences, 2016, 105(2): 519-529.
20	Gerhardt A, McGraw N R, Schwartz D K, et al. Protein aggregation and particle formation in prefilled glass syringes[J]. Journal of Pharmaceutical Sciences, 2014, 103(6): 1601-1612.
21	Qi L, Liu J, Ronk M, et al. A holistic approach of extractables and leachables assessment of rubber stoppered glass vial systems for biotechnology products[J]. Journal of Pharmaceutical Sciences, 2021, 110(11): 3580-3593.
22	Oom A, Poggi M, Wikström J, et al. Surface interactions of monoclonal antibodies characterized by quartz crystal microbalance with dissipation: impact of hydrophobicity and protein self-interactions[J]. Journal of Pharmaceutical Sciences, 2012, 101(2): 519-529.
23	Gao F F. Adsorption of mussel protein on polymer antifouling membranes: a molecular dynamics study[J]. Molecules (Basel, Switzerland), 2021, 26(18): 5660.
24	Marsh R J, Jones R A L, Sferrazza M. Adsorption and displacement of a globular protein on hydrophilic and hydrophobic surfaces[J]. Colloids and Surfaces B: Biointerfaces, 2002, 23(1): 31-42.
25	Lefebvre G, Maze A, Jimenez R A P, et al. Surfactant protection efficacy at surfaces varies with the nature of hydrophobic materials[J]. Pharmaceutical Research, 2021, 38(12): 2157-2166.
26	Bainor A, Chang L, McQuade T J, et al. Bicinchoninic acid (BCA) assay in low volume[J]. Analytical Biochemistry, 2011, 410(2): 310-312.
27	Gerhardt A, Bonam K, Bee J S, et al. Ionic strength affects tertiary structure and aggregation propensity of a monoclonal antibody adsorbed to silicone oil-water interfaces[J]. Journal of Pharmaceutical Sciences, 2013, 102(2): 429-440.
28	Hawe A, Sutter M, Jiskoot W. Extrinsic fluorescent dyes as tools for protein characterization[J]. Pharmaceutical Research, 2008, 25(7): 1487-1499.
29	Ying P Q, Yu Y, Jin G, et al. Competitive protein adsorption studied with atomic force microscopy and imaging ellipsometry[J]. Colloids and Surfaces B: Biointerfaces, 2003, 32(1): 1-10.
30	Rabe M, Verdes D, Seeger S. Understanding protein adsorption phenomena at solid surfaces[J]. Advances in Colloid and Interface Science, 2011, 162(1/2): 87-106.
31	Chen S F, Liu L Y, Zhou J, et al. Controlling antibody orientation on charged self-assembled monolayers[J]. Langmuir, 2003, 19: 2859-2864.
32	Bee J S, Schwartz D K, Trabelsi S, et al. Production of particles of therapeutic proteins at the air-water interface during compression/dilation cycles[J]. Soft Matter, 2012, 8(40): 10329-10335.
33	Haynes Charles A, Willem N. Globular proteins at solid/liquid interfaces[J]. Colloids and Surfaces B: Biointerfaces, 1994, 2(6): 517-566.
34	Mathes J, Friess W. Influence of pH and ionic strength on IgG adsorption to vials[J]. European Journal of Pharmaceutics and Biopharmaceutics, 2011, 78(2): 239-247.
35	Quan X B, Liu J, Zhou J. Multiscale modeling and simulations of protein adsorption: progresses and perspectives[J]. Current Opinion in Colloid & Interface Science, 2019, 41: 74-85.
36	Wu H, Randolph T W. Rapid quantification of protein particles in high-concentration antibody formulations[J]. Journal of Pharmaceutical Sciences, 2019, 108(3): 1110-1116.

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