Many companies, including Strike Pharma, are developing biologicals for individualized immunotherapeutic cancer treatments. The possibility to combine a bispecific antibody with a myriad of endogenous antigenic peptides opens the doors for highly personalized therapies. Setting up and using analytical assays is key to evaluate aggregation and overcome aggregation patterns of biologicals during CMC development. The aim of this project was to assess size exclusion chromatography (SEC) as an analytical method and subsequently evaluate several drug formulations that could be suitable for subcutaneous administration of a peptide and antibody conjugate mix. The formulations were based on a 25 mM histidine buffer pH 6.0, that had been optimized for the antibody alone, with different additives. By utilizing SEC coupled to UV-detection at 280 nm, aggregates were detected and quantified. The most effective excipients were dimethyl sulfoxide, polyethylene glycol 400 and arginine. Two different peptide-tags were compared and the pTag9mer-mut2 variant was more favorable than pTag9mer-mut1 in limiting aggregate formation with highest success rates at 1.5 mg/mL protein concentrations and the fulfillment of the high molecular weight ≤ 5% criterion. Combining antibody and peptide containing pTag9mer-mut1 in a pH 9.0 histidine buffer with added arginine engendered the least aggregates compared to any pH 6.0 formulation. However, the instability of the antibody in pH 9.0 and the risk of deamidation makes this less suitable. Future considerations include changing the administration method or using pump injection strategy, which allows higher injection volumes to limit aggregation by lowering protein concentrations.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:uu-533886 |
| Date | January 2024 |
| Creators | Imedashvili, Sumay |
| Publisher | Uppsala universitet, Institutionen för farmaci |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | UPTEC K, 1650-8297 ; 24022 |
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