Sorbitol Crystallization-Induced Aggregation in Frozen mAb Formulations

Piedmonte, Deirdre Murphy; Hair, Alison; Baker, Priti; Brych, Lejla; Nagapudi, Karthik; Lin, Hong; Cao, Wenjin; Hershenson, Susan; Ratnaswamy, Gayathri

doi:10.1002/jps.24141

Cited by 15 publications

(3 citation statements)

References 47 publications

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“…Sucrose was preferred over trehalose, mannitol, and sorbitol (even though all 3 showed similar or slightly better thermal stability profiles compared to sucrose) because of cost or the known tendency of these additives to crystallize out during freezing and thawing. [31][32][33] In addition, equal amounts (% w/v) of the 2 polyols (vs. sucrose or trehalose) impart higher solution osmolality which is less desirable for parenteral administration due to their hypertonic nature. 34,35 Evaluation of Candidate Formulations for Bulk Drug Substance of Three NRRV Antigens Eight candidate formulations (F2-F9) listed in Table 2 were designed based on the aforementioned results.…”

Section: Evaluating Stabilizing Excipients To Minimize Shaking-inducementioning

confidence: 99%

Characterizing and Minimizing Aggregation and Particle Formation of Three Recombinant Fusion-Protein Bulk Antigens for Use in a Candidate Trivalent Rotavirus Vaccine

Sahni

Hickey

Robertson

et al. 2020

Journal of Pharmaceutical Sciences

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In a companion paper, the structural integrity, conformational stability, and degradation mechanisms of 3 recombinant fusion-protein antigens comprising a non-replicating rotavirus (NRRV) vaccine candidate (currently being evaluated in early-stage clinical trials) are described. In this work, we focus on the aggregation propensity of the 3 NRRV antigens coupled to formulation development studies to identify common frozen bulk candidate formulations. The P2-VP8-P[8] antigen was most susceptible to shaking and freeze-thaweinduced aggregation and particle formation. Each NRRV antigen formed aggregates with structurally altered protein (with exposed apolar regions and intermolecular b-sheet) and dimers containing a non-native disulfide bond. From excipient screening studies with P2-VP8-P[8], sugars or polyols (e.g., sucrose, trehalose, mannitol, sorbitol) and various detergents (e.g., Pluronic F-68, polysorbate 20 and 80, PEG-3350) were identified as stabilizers against aggregation. By combining promising additives, candidate bulk formulations were optimized to not only minimize agitation-induced aggregation, but also particle formation due to freeze-thaw stress of P2-VP8-P[8] antigen. Owing to limited material availability, stabilization of the P2-VP8-P[4] and P2-VP8-P[6] was confirmed with the lead candidate P2-VP8-P[8] formulations. The optimization of these bulk NRRV candidate formulations is discussed in the context of subsequent drug product formulations in the presence of aluminum adjuvants.

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Section: Evaluating Stabilizing Excipients To Minimize Shaking-inducementioning

confidence: 99%

Characterizing and Minimizing Aggregation and Particle Formation of Three Recombinant Fusion-Protein Bulk Antigens for Use in a Candidate Trivalent Rotavirus Vaccine

Sahni

Hickey

Robertson

et al. 2020

Journal of Pharmaceutical Sciences

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“…Similarly, sorbitol and lactose solutions also showed higher number of sub-visible particle formation than sucrose solutions during agitation. Moreover, sorbitol and mannitol may manifest a lesser ability to protect during freezing and thawing compared to disaccharides at equivalent solution osmolality values [18] , [19] , [20] , [21] , a consideration of importance for dmLT as described below. Finally, lactose is a reducing sugar which can glycate Lys residues in proteins (as was observed with dmLT, Toprani et al, 2017, submitted), a reaction that can potentially lead to protein instability as well as formation of advanced glycation products [22] , [23] .…”

Section: Resultsmentioning

confidence: 99%

Development of a candidate stabilizing formulation for bulk storage of a double mutant heat labile toxin (dmLT) protein based adjuvant

Toprani

Sahni

Hickey

et al. 2017

Vaccine

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This work describes the formulation design and development of a novel protein based adjuvant, a double mutant of heat labile toxin (dmLT), based on knowledge of the protein’s structural integrity and physicochemical degradation pathways. Various classes of pharmaceutical excipients were screened for their stabilizing effect on dmLT during exposure to thermal and agitation stresses as monitored by high throughput analytical assays for dmLT degradation. Sucrose, phosphate, sodium chloride, methionine and polysorbate-80 were identified as potential stabilizers that protected dmLT against either conformational destabilization, aggregation/particle formation or chemical degradation (e.g., Met oxidation and Lys glycation). Different combinations and concentrations of the selected stabilizers were then evaluated to further optimize dmLT stability while maintaining pharmaceutically acceptable ranges of solution pH and osmolality. The effect of multiple freeze-thaw (FT) cycles on the physical stability of candidate bulk formulations was also examined. Increasing the polysorbate-80 concentration to 0.1% in the lead candidate bulk formulation mitigated the loss of protein mass during FT. This formulation development study enabled the design of a new bulk formulation of the dmLT adjuvant and provides flexibility for future use in combination with a variety of different vaccine dosage forms with different antigens.

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“…In the literature, SOR was used to stabilize mAb in solution or during a freeze-drying process. However, aggregation was promoted due to the recrystallization of SOR during storage (Kamerzell et al, 2011;Piedmonte et al, 2015). Crystallization may lead to a loss of proteinstabilizer interactions .…”

Section: Tablementioning

confidence: 99%

Development of mAb-loaded 3D-printed (FDM) implantable devices based on PLGA

Carlier

Marquette

Peerboom

et al. 2021

International Journal of Pharmaceutics

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The main objective of this work was to explore the feasibility to print monoclonal antibody (mAb)-loaded implantable systems using fused-deposition modelling (FDM) to build complex dosage form designs. Indeed, to our knowledge, this work is the first investigation of mAb-loaded devices using FDM. To make this possible, different steps were developed and optimized. A mAb solution was stabilized using trehalose (TRE), sucrose (SUC), hydroxypropyl-β-cyclodextrin (HP-β-CD), sorbitol or inulin (INU) in order to be spray dried (SD). Printable filaments were then made of poly(lactide-co-glycolide) (PLGA) and mAb powder (15% w/w) using hot melt extrusion (HME). The FDM process was optimized to print these filaments without altering the mAb stability. TRE was selected and associated to L-leucine (LEU) to increase the mAb stability. The stability was then evaluated considering high and low molecular weight species levels. The mAb-based devices were well-stabilized with the selected excipients during both the HME and the FDM processes. The 3D-printed devices showed sustained-release profiles with a low burst effect. The mAb-binding capacity was preserved up to 70% following the whole fabrication process. These promising results demonstrate that FDM could be used to produce mAbloaded devices with good stability, affinity and sustained-release profiles of the mAb.

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Sorbitol Crystallization-Induced Aggregation in Frozen mAb Formulations

Cited by 15 publications

References 47 publications

Characterizing and Minimizing Aggregation and Particle Formation of Three Recombinant Fusion-Protein Bulk Antigens for Use in a Candidate Trivalent Rotavirus Vaccine

Characterizing and Minimizing Aggregation and Particle Formation of Three Recombinant Fusion-Protein Bulk Antigens for Use in a Candidate Trivalent Rotavirus Vaccine

Development of a candidate stabilizing formulation for bulk storage of a double mutant heat labile toxin (dmLT) protein based adjuvant

Development of mAb-loaded 3D-printed (FDM) implantable devices based on PLGA

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