title: Impact of controlled ice nucleation on process performance and quality attributes of a lyophilized monoclonal antibody.
abstract: An efficient and potentially scalable technology was evaluated to control the ice nucleation step of the freezing process for a model monoclonal antibody formulation and the effect on process performance and quality attributes of the final lyophilized product was compared with the conventional shelf ramping method of freezing. Controlled ice nucleation resulted in uniform nucleation at temperatures between -2.3 and -3.2 °C while uncontrolled nucleation resulted in random nucleation at temperatures between -10 and -16.4 °C. The sublimation rate (dm/dt) during primary drying was higher in the controlled nucleation cycle (0.13 g/h/vial) than in the uncontrolled nucleation cycle (0.11 g/h/vial). This was due to the formation of larger ice crystals, leading to lower product resistance (Rp) and 19% reduction in the primary drying for the controlled nucleation cycle. Controlled ice nucleation resulted in lyophilized cakes with more acceptable appearance, no visible collapse or shrinkage and decreased reconstitution times compared with uncontrolled nucleation. There were no observed differences in the particle size, concentration (A280 nm) and presence of aggregates (A410 nm) between the two nucleation cycles when the lyophilized cakes were reconstituted. These were confirmed by SEC and protein A-HPLC analyses which showed similar peak shapes and retention times between the two cycles. However, uncontrolled nucleation resulted in cakes with larger specific surface area (0.90 m(2)/g) than controlled nucleation (0.46 m(2)/g). SEM images of the lyophilized cakes from uncontrolled nucleation revealed a sponge-like morphology with smaller pores while cakes from controlled nucleation cycle revealed plate-like structures with more open and larger pores. While controlled nucleation resulted in a final product with a higher residual moisture content (2.1±0.08%) than uncontrolled nucleation (1.62±0.11%), this was resolved by increasing the secondary drying temperature.
authors.affiliation: Division of Product Quality Research, Office of Testing and Research, OPS, CDER, FDA, Rockville, MD, USA.
authors.forename: David
authors.initials: D
authors.lastname: Awotwe-Otoo
authors.forename: Cyrus
authors.initials: C
authors.lastname: Agarabi
authors.forename: Erik K
authors.initials: EK
authors.lastname: Read
authors.forename: Scott
authors.initials: S
authors.lastname: Lute
authors.forename: Kurt A
authors.initials: KA
authors.lastname: Brorson
authors.forename: Mansoor A
authors.initials: MA
authors.lastname: Khan
authors.forename: Rakhi B
authors.initials: RB
authors.lastname: Shah
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical; D004339:Drug Compounding / Q000379:methods*; D005612:Freeze Drying; D007074:Immunoglobulin G / Q000737:chemistry*; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains