Formulation mitigations for particle formation induced by enzymatic hydrolysis of polysorbate 20 in protein-based drug products: insights from a full-factorial longitudinal study

AAPS Open

Table 2 Effect estimates for three formulation parameters and their interactions on particle formation and PS20 degradation. The formulation parameters—[mAb], initial [PS20], and pH—were tested in a DP stability study at 5 °C with a full-factorial design. Particle formation was evaluated as duration with no VPs (by the Ph. Eur./USP method) and duration with low SVPs (< 350 counts for ≥ 2 µm particles/mL by HIAC liquid particle counter). PS20 degradation was quantified by measuring [PS20] in % w/v by HPLC-ELSD (Fig. 6) and [LA] in µg/mL by LC–MS (Fig. 8) over DP storage. The rate of PS20 change (d[PS20]/dt; Fig. 7) and rate of LA change (d[LA]/dt; Fig. 9) were calculated using the derivatives from fitting to a longitudinal model

Formulation parameters				Effect estimates (midpoint to high end of range)
Formulation parameters				Duration (weeks)		Rate (week⁻¹)
Parameter(s) (units)		Term	Coded range	No VPs	Low SVPs	d[PS20]/dt(× 10⁻⁴)	d[LA]/dt
Main effects	mAb concentration (mg/mL)	[mAb]	10–40	− 5.95^*	− 6.78^*	− 2.39^*	0.41^*
	Initial PS20 concentration (% w/v)	Initial [PS20]	0.02–0.06	3.88^*	4.19^*	− 1.64^*	0.26^*
	pH	pH	5.0–6.0	0.50	0.38	− 0.31^*	0.10^*
Interactions	mAb concentration (mg/mL) and initial PS20 concentration (% w/v)	[mAb] × initial [PS20]	10–40; 0.02–0.06	− 1.88^*	− 2.44^*	− 1.35^*	0.16^*
	mAb concentration (mg/mL) and pH	[mAb] × pH	10–40; 5.0–6.0	− 0.50	0.13	0.12	0.02
	Initial PS20 concentration (% w/v) and pH	Initial [PS20] × pH	0.02–0.06; 5.0–6.0	0.25	0.19	− 0.24^*	0.04
	mAb concentration (mg/mL), initial PS20 concentration (% w/v), and pH	[mAb] × initial [PS20] × pH	10–40; 0.02–0.06; 5.0–6.0	0.15	0.41	− 0.18	0.03