Characterization of the Physical Stability of a Lyophilized IgG1 mAb after Accelerated Shipping-Like Stress

Abstract

Upon exposure to shaking stress, an IgG1 mAb formulation in both liquid and lyophilized state formed subvisible particles. Since freeze-drying is expected to minimize protein physical instability under these conditions, the extent and nature of aggregate formation in the lyophilized preparation was examined using a variety of particle characterization techniques. The effect of formulation variables such as residual moisture content, reconstitution rate, and reconstitution medium were examined. Upon reconstitution of shake-stressed lyophilized mAb, differences in protein particle size and number were observed by Microflow Digital Imaging (MFI), with the reconstitution medium having the largest impact. Shake-stress had minor effects on the structure of protein within the particles as shown by SDS-PAGE and FTIR analysis. The lyophilized mAb was shake-stressed to different extents and stored for 3 months at different temperatures. Both extent of cake collapse and storage temperature affected the physical stability of the shake-stressed lyophilized mAb upon subsequent storage. These findings demonstrate that physical degradation upon shaking of a lyophilized IgG1 mAb formulation includes not only cake breakage, but also results in an increase in subvisible particles and turbidity upon reconstitution. The shaking-induced cake breakage of the lyophilized IgG1 mAb formulation also resulted in decreased physical stability upon storage.

Assessing the Utility of Circular Dichroism and FTIR Spectroscopy in Monoclonal-Antibody Comparability Studies

“…Studies with IgG1 mAbs indicate that changes in higher order structure are sometimes detected by CD and/or FTIR spectroscopy in product samples that underwent extreme stress not typically encountered during routine manufacturing [e.g., incubation at high temperature (90°C) and extreme agitation of lyophilized products], whereas in other cases, contrary conclusions were made after significant H 2 O 2 oxidation or acid exposure . In this account, we outline and conclude that CD and FTIR are insufficiently sensitive to find measurable higher order structure changes in samples after stressed to a degree that is representative of current manufacturing processes.…”

An overview of lyophilization: troubleshooting the challenges and pharmaceutical applications

“…Higher cracking was seen in 10R vials than 2R vials with constant fill volume [25]. Srivalli Telikepalli studied that shipping stress can play a major role in breakage of the cake [27]. To prevent this defect product temperature can be kept lower in drying step and cycle optimization should be done with trial-and-error method.…”

Mechanical Shock During Shipping of Medications: The Roles of Packaging and Transportation Vendors

“…[2][3][4] Mechanical shock has also been shown to have deleterious effects on therapeutic protein molecules, with shock in the form of drop stress causing particle formation and protein aggregation as well as potential damage to the autoinjector or syringe. [5][6][7][8][9] Experience with recombinant therapeutic proteins and plasmaderived medications (e.g., intravenous immunoglobulin) has established that these types of particles and protein aggregates may result in immunologic consequences including hypersensitivity reactions or production of anti-therapeutic antibodies. 9 The end effects of these types of reactions include direct harm in the form of adverse drug reaction or secondary issues related to loss in efficacy of the drug product, with resultant flare of underlying disease.…”