Use of Transient Transfection for cGMP Manufacturing of eOD-GT8 60mer, a Self-Assembling Nanoparticle Germline-Targeting HIV-1 Vaccine Candidate
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pharmaceutics-16-00742-v2.pdf
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2.46 MB
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Author(s)
Sharma, Vaneet K.
Menis, Sergey
Brower, Evan T.
Sayeed, Eddy
Ackland, Jim
Lombardo, Angela
Cottrell, Christopher A.
Torres, Jonathan L.
Hassell, Thomas
Ward, Andrew B.
Date Issued
May 30, 2024
Journal
Pharmaceutics
Publisher
MDPI AG
Citation
Sharma, V.K.; Menis, S.; Brower, E.T.; Sayeed, E.; Ackland, J.; Lombardo, A.; Cottrell, C.A.; Torres, J.L.; Hassell, T.; Ward, A.B.; et al. Use of Transient Transfection for cGMP Manufacturing of eOD-GT8 60mer, a Self-Assembling Nanoparticle Germline-Targeting HIV-1 Vaccine Candidate. Pharmaceutics 2024, 16, 742.
Version
Final published version
Abstract
We describe the current Good Manufacturing Practice (cGMP) production and subsequent characterization of eOD-GT8 60mer, a glycosylated self-assembling nanoparticle HIV-1 vaccine candidate and germline targeting priming immunogen. Production was carried out via transient expression in the human embryonic kidney 293 (HEK293) cell line followed by a combination of purification techniques. A large-scale cGMP (200 L) production run yielded 354 mg of the purified eOD-GT8 60mer drug product material, which was formulated at 1 mg/mL in 10% sucrose in phosphate-buffered saline (PBS) at pH 7.2. The clinical trial material was comprehensively characterized for purity, antigenicity, glycan composition, amino acid sequence, and aggregation and by several safety-related tests during cGMP lot release. A comparison of the purified products produced at the 1 L scale and 200 L cGMP scale demonstrated the consistency and robustness of the transient transfection upstream process and the downstream purification strategies. The cGMP clinical trial material was tested in a Phase 1 clinical trial (NCT03547245), is currently being stored at −80 °C, and is on a stability testing program as per regulatory guidelines. The methods described here illustrate the utility of transient transfection for cGMP production of complex products such as glycosylated self-assembling nanoparticles.
MIT Department
Ragon Institute of MGH, MIT and Harvard
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DOI of Published Version
https://doi.org/10.3390/pharmaceutics16060742
