Characterization of critical quality attributes (CQAs) for bispecific antigen- binding biotherapeutic (BABB) through comprehensive analysis using separation techniques and footprinting approaches coupled with mass spectrometry

Permanent URL: http://hdl.handle.net/2047/D20537505
Title:
Characterization of critical quality attributes (CQAs) for bispecific antigen- binding biotherapeutic (BABB) through comprehensive analysis using separation techniques and footprinting approaches coupled with mass spectrometry
Creator:
Shah, Arnik (Author)
Contributor:
Ivanov, Alexander R (Thesis advisor)
Agar, Jeffrey (Committee member)
Zhenyu, Tian (Committee member)
Cui, Weidong (Committee member)
Thiel, Tiffany (Committee member)
Language:
English
Publisher:
Boston, Massachusetts : Northeastern University, 2023
Date Accepted:
June 2023
Date Awarded:
August 2023
Type of resource:
Text
Genre:
Dissertations
Format:
electronic
Digital origin:
born digital
Abstract/Description:
Bispecific antigen-binding biotherapeutics (BABB) are a novel class of therapeutics developed to combat various disease conditions. Identifying critical quality attributes (CQAs) for novel therapeutics is important in establishing quality by design (QbD) to achieve consistent product quality to ensure safety and efficacy to patients. BABB molecules are produced at low concentrations due to their high potency compared to traditional monoclonal antibodies. This creates challenges in characterizing the product based on traditional analytical approaches. Mass spectrometry-based methods have recently emerged as powerful analytical tools to characterize post-translation modifications (PTMs) of biotherapeutics. In this work, we explored different separation techniques and footprinting approaches coupled with mass spectrometry to identify critical quality attributes of BABB molecules. In the first chapter, we describe how we utilized microfluidic-based capillary zone electrophoresis coupled with mass spectrometry to identify fragmentation sites of this novel therapeutic. In the second chapter, we overview how native mass spectrometry-based workflow was developed to characterize modifications present on the BABB molecule, offering rapid and unbiased characterization of predominant and low abundance PTMs compared to more traditional approaches. In the third chapter, we explore applications of covalent labeling and mass spectrometry to identify subtle changes in the BABB molecule under two states (native vs. stressed) that can impact the product quality. These workflows provide in-depth characterization of PTMs and changes in conformation at the amino acid level for the BABB molecule.Application of such workflows at different stages of drug development can aid in the rapid and unbiased characterization of CQAs related to BABB and other novel modalities of protein therapeutics. --Author's abstract
Subjects and keywords:
attribute monitoring
Biologics-characterization
Chromatography/capillary electrophoretic separationa
covalent labeling
Mass spectrometry
Native mass spectrometry
Analytical chemistry
Biochemistry
Chemistry
DOI:
https://doi.org/10.17760/D20537505
Permanent URL:
http://hdl.handle.net/2047/D20537505
Use and reproduction:
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