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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 the liquid and lyophilized state formed subvisible particles. Because freeze-drying was expected to minimize protein physical instability under these conditions, the extent and nature of aggregate formation in the lyophilized preparation were examined using a variety of particle characterization techniques. The effects of formulation variables such as residual moisture content, reconstitution rate, and reconstitution medium were also examined. Upon reconstitution of shake-stressed lyophilized mAb, differences in protein particle size and number were observed by microflow digital imaging, 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 shake-induced cake breakage of the lyophilized IgG1 mAb formulation also resulted in decreased physical stability upon storage.

SUBMITTER: Telikepalli S

PROVIDER: S-EPMC4312241 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Characterization of the physical stability of a lyophilized IgG1 mAb after accelerated shipping-like stress.

Telikepalli Srivalli S Kumru Ozan S OS Kim Jae Hyun JH Joshi Sangeeta B SB O'Berry Kristin B KB Blake-Haskins Angela W AW Perkins Melissa D MD Middaugh C Russell CR Volkin David B DB

Journal of pharmaceutical sciences 20141217 2

Upon exposure to shaking stress, an IgG1 mAb formulation in both the liquid and lyophilized state formed subvisible particles. Because freeze-drying was expected to minimize protein physical instability under these conditions, the extent and nature of aggregate formation in the lyophilized preparation were examined using a variety of particle characterization techniques. The effects of formulation variables such as residual moisture content, reconstitution rate, and reconstitution medium were al ...[more]

PMID: 25522000

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Project description:Some concerns have been raised recently about the assay of corticosterone vs. cortisol in poultry species. Thus, we tested the hypothesis that ducks secrete both glucocorticoids. First, we validated two commercially ELISA kits for the two glucocorticoids by first charcoal stripping duck serum in order to remove all steroid hormones. We ran serial dilutions of spiked, charcoal-stripped serum on kits of opposite glucocorticoid as well as a serial dilution using the respective ELISA buffer of the opposite assay kit. We found that the glucocorticoid standard curve in duck serum matched the respective curve in that kit's own buffer. However, when the opposite hormone was run in each kit in both duck serum or ELISA buffer, a near zero slope was obtained. Second, we further validated the presence of both glucocorticoids using mass spectrometry. Third, we tested the hypothesis that exogenous ACTH would stimulate the release of both corticosterone and cortisol. And, fourth, we tested the hypothesis that each glucocorticoid would have different serum levels in response to shipping stress. To test this hypothesis, we collected serum from 10 drakes and 10 hens from 2 flocks (N = 20 per time point per sex): 24 h prior to shipping, at shipping as ducks were walked off the truck, 24 h after shipping, and 1 wk after shipping. Data were analyzed by 2-way repeated measures ANOVA. Surprisingly, we also observed a sex difference in both glucocorticoid levels in that hens showed higher (P < 0.01) serum levels than did drakes at all-time points in response to either ACTH or transportation. Finally, no differences were observed in either glucocorticoid levels associated with shipping in either sex. The fact that both glucocorticoids are released in measurable amounts lends to the possibility that they may be differentially regulated, or at least there is a sex difference in the neural pathways associated with glucocorticoid release in ducks. Although corticosterone is the likely predominate glucocorticoid in ducks, serious attention should be given to the role of cortisol in poultry. Further consideration of sex, age, and timing of blood collection to stressor needs to be considered when assessing glucocorticoid levels in any avian species.

Dataset Information

Characterization of the physical stability of a lyophilized IgG1 mAb after accelerated shipping-like stress.

Publications

Characterization of the physical stability of a lyophilized IgG1 mAb after accelerated shipping-like stress.

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