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The Effects of Excipients on Freeze-dried Monoclonal Antibody Formulation Degradation and Sub-Visible Particle Formation during Shaking

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Abstract

Purposes

We previously reported an unexpected phenomenon that shaking stress could cause more protein degradation in freeze-dried monoclonal antibody (mAb) formulations than liquid ones (J Pharm Sci, 2022, 2134). The main purposes of the present study were to investigate the effects of shaking stress on protein degradation and sub-visible particle (SbVP) formation in freeze-dried mAb formulations, and to analyze the factors influencing protein degradation during production and transportation.

Methods

The aggregation behavior of mAb-X formulations during production and transportation was simulated by shaking at a rate of 300 rpm at 25°C for 24 h. The contents of particles and monomers were analyzed by micro-flow imaging, dynamic light scattering, size exclusion chromatography, and ultraviolet − visible (UV–Vis) spectroscopy to compare the protective effects of excipients on the aggregation of mAb-X.

Results

Shaking stress could cause protein degradation in freeze-dried mAb-X formulations, while surfactant, appropriate pH, polyol mannitol, and high protein concentration could impact SbVP generation. Water content had little effect on freeze-dried protein degradation during shaking, as far as the water content was controlled in the acceptable range as recommended by mainstream pharmacopoeias (i.e., less than 3%).

Conclusions

Shaking stress can reduce the physical stability of freeze-dried mAb formulations, and the addition of surfactants, polyol mannitol, and a high protein concentration have protective effects against the degradation of model mAb formulations induced by shaking stress. The experimental results provide new insight for the development of freeze-dried mAb formulations.

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Data Availability

The data will be available upon request.

Abbreviations

DLS:

Dynamic light scattering

HES:

Hydroxyethyl starch

His-HCl:

Histidine-HCl

kD :

Diffusion interaction parameter

mAb:

Monoclonal antibody

MFI:

Micro-flow imaging

NMPA:

National Medicinal Products Administration of China

PDI:

Distribution coefficient

PES:

Polyethersulfone

PS:

Polysorbate

SbVP:

Sub-visible particle

SE − HPLC:

Size exclusion high-performance liquid chromatography

UV − Vis:

Ultraviolet − visible spectroscopy

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Acknowledgements

This study was supported by the National Nature Science Foundation of China (Grant No. 81741144) and the Huadong Medicine Joint Funds of the Zhejiang Provincial Natural Science Foundation of China (Grant No. LHDMZ24H300003). We thank Ms. Haihong Hu for laboratory management.

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Authors and Affiliations

  1. Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China

    Meng-Jia Jin, Xin-Zhe Ge, Qiong Huang, Jia-Wei Liu & Wei-Jie Fang

  2. Hangzhou Institute of Innovative Medicine, Zhejiang University, Hangzhou, 310016, China

    Meng-Jia Jin, Xin-Zhe Ge, Qiong Huang, Jia-Wei Liu & Wei-Jie Fang

  3. Datta Meghe College of Pharmacy, Datta Meghe Institute of Higher Education & Research, Sawangi, Wardha, India

    Rahul G. Ingle

  4. Zhejiang Bioray Biopharmaceutical Co., Taizhou, 317000, China

    Dong Gao

  5. Innovation Center of Translational Pharmacy, Jinhua Institute of Zhejiang University, Jinhua, 321000, China

    Wei-Jie Fang

  6. Taizhou Institute of Zhejiang University, Taizhou, 317000, China

    Wei-Jie Fang

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  1. Meng-Jia Jin
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Jin, MJ., Ge, XZ., Huang, Q. et al. The Effects of Excipients on Freeze-dried Monoclonal Antibody Formulation Degradation and Sub-Visible Particle Formation during Shaking. Pharm Res 41, 321–334 (2024). https://doi.org/10.1007/s11095-024-03657-7

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