Light-assisted drying (LAD) for anhydrous preservation of biologics: processing sample volumes comparable to a therapeutic dose

Search for this publication on Google Scholar
Furr, D. (2022). Light-assisted drying (LAD) for anhydrous preservation of biologics: processing sample volumes comparable to a therapeutic dose. Unc Charlotte Electronic Theses And Dissertations.
Download PDF

Analytics

34 views ◎
17 downloads ⇓


Abstract

Protein-based products have been developed to treat a range of conditions and are used in vaccines and assays. A challenge in the development of these products is maintaining the protein in the folded state during processing and storage. The most common method of stabilizing proteins for storage is lyophilization (freeze drying). However, this process remains expensive and many proteins that are lyophilized must be refrigerated or frozen to maintain functionality. Cold-chain storage can be challenging and expensive for the transportation and storage of biologics, especially in low-resource settings. Recent research has demonstrated that anhydrous preservation in a trehalose amorphous solid matrix offers an alternative to freeze drying for the preservation of biologics. We have previously described a new processing technique, light assisted drying (LAD), to create trehalose preservation matrices of small volume (40 μL) samples. LAD uses illumination by near-infrared laser light to selectively heat water and speed dehydration. In this study we apply the LAD technique to large volume samples (250 μL) that are more comparable to therapeutic doses. A model protein, lysozyme, was LAD processed then stored for 1 month. The end moisture content of samples was determined immediately after processing and then again after storage. The thermal histories of samples were monitored during processing to determine the optimal drying time. The trehalose matrix was characterized using polarized light imaging to determine if crystallization occurred during storage, damaging embedded proteins. Raman spectroscopy was used to determine the distribution of water content. Karl-Fischer (KF) Titration was used to analyze the actual water content of the samples. These studies indicate that LAD can effectively stabilize large volume samples.

Details
Author

Furr, Daniel

Title

Light-assisted drying (LAD) for anhydrous preservation of biologics: processing sample volumes comparable to a therapeutic dose

Physical Description

1 online resource (54 pages) : PDF

Date

2022

Degree Granting Institution

University of North Carolina at Charlotte

Abstract

Protein-based products have been developed to treat a range of conditions and are used in vaccines and assays. A challenge in the development of these products is maintaining the protein in the folded state during processing and storage. The most common method of stabilizing proteins for storage is lyophilization (freeze drying). However, this process remains expensive and many proteins that are lyophilized must be refrigerated or frozen to maintain functionality. Cold-chain storage can be challenging and expensive for the transportation and storage of biologics, especially in low-resource settings. Recent research has demonstrated that anhydrous preservation in a trehalose amorphous solid matrix offers an alternative to freeze drying for the preservation of biologics. We have previously described a new processing technique, light assisted drying (LAD), to create trehalose preservation matrices of small volume (40 μL) samples. LAD uses illumination by near-infrared laser light to selectively heat water and speed dehydration. In this study we apply the LAD technique to large volume samples (250 μL) that are more comparable to therapeutic doses. A model protein, lysozyme, was LAD processed then stored for 1 month. The end moisture content of samples was determined immediately after processing and then again after storage. The thermal histories of samples were monitored during processing to determine the optimal drying time. The trehalose matrix was characterized using polarized light imaging to determine if crystallization occurred during storage, damaging embedded proteins. Raman spectroscopy was used to determine the distribution of water content. Karl-Fischer (KF) Titration was used to analyze the actual water content of the samples. These studies indicate that LAD can effectively stabilize large volume samples.

Genre

masters theses

Subjects--Topics

Physics

Degree

M.S.

Keywords

Biopreservation
Lad
Protein Storage
Trehalose

Subject Area

Optical Science & Engineering

Advisor(s)

Trammell, Susan

Committee Members

Jacobs, Donald
Zhang, Yong

Degree Note

Thesis (M.S.)--University of North Carolina at Charlotte, 2022.

Rights Statement

This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). For additional information, see http://rightsstatements.org/page/InC/1.0/.

Rights Holder Information

Copyright is held by the author unless otherwise indicated.

Identifier

Furr_uncc_0694N_13304

Permalink

http://hdl.handle.net/20.500.13093/etd:3102