Use of Transient Transfection for cGMP Manufacturing of eOD-GT8 60mer, a Self-Assembling Nanoparticle Germline-Targeting HIV-1 Vaccine Candidate.
Vaneet K SharmaSergey MenisEvan T BrowerEddy SayeedJim AcklandAngela LombardoChristopher A CottrellJonathan L TorresThomas HassellAndrew B WardVadim TsvetnitskyWilliam R SchiefPublished in: Pharmaceutics (2024)
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.
Keyphrases
- nitric oxide
- clinical trial
- protein kinase
- cerebral ischemia
- antiretroviral therapy
- amino acid
- phase ii
- hiv infected
- hiv positive
- open label
- human immunodeficiency virus
- healthcare
- hepatitis c virus
- endothelial cells
- hiv testing
- quality improvement
- cancer therapy
- drug delivery
- brain injury
- dna damage
- binding protein
- ionic liquid
- electronic health record
- adverse drug
- south africa