Strategy for Generating Sequence-Defined Aptamer Reagent Sets for Detecting Protein Contaminants in Biotherapeutics

Strategy for Generating Sequence-Defined Aptamer Reagent Sets for Detecting Protein Contaminants in Biotherapeutics

Biologic drugs are typically manufactured in mammalian host cells, and it is critical from a drug safety and efficacy perspective to detect and remove host cell proteins (HCPs) during production. This is currently achieved with sets of polyclonal antibodies (pAbs), but these suffer from critical sho...

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Bibliographic Details
Published in:Analytical chemistry (Washington) Vol. 90; no. 5; pp. 3262 - 3269
Main Authors: McGivney, James B, Csordas, Andrew T, Walker, Faye M, Bagley, Elizabeth R, Gruber, Emily M, Mage, Peter L, Casas-Finet, Jose, Nakamoto, Margaret A, Eisenstein, Michael, Larkin, Christopher J, Strouse, Robert J, Soh, H. Tom
Format: Journal Article
Language:English
Published: United States American Chemical Society 06-03-2018
Subjects:
Analytical chemistry
Antibodies
Aptamers
Cell lines
Cells
Chemistry
Contaminants
Deoxyribonucleic acid
DNA
Drugs
High-throughput screening
Immunoglobulins
Isoelectric points
Pharmacovigilance
Polyclonal antibodies
Proteins
Reagents
Strategy
Toxicity
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Summary:Biologic drugs are typically manufactured in mammalian host cells, and it is critical from a drug safety and efficacy perspective to detect and remove host cell proteins (HCPs) during production. This is currently achieved with sets of polyclonal antibodies (pAbs), but these suffer from critical shortcomings because their composition is inherently undefined, and they cannot detect nonimmunogenic HCPs. In this work, we report a high-throughput screening and array-based binding characterization strategy that we employed to generate a set of aptamers that overcomes these limitations to achieve sensitive, broad-spectrum detection of HCPs from the widely used Chinese hamster ovary (CHO) cell line. We identified a set of 32 DNA aptamers that achieve better sensitivity than a commercial pAb reagent set and can detect a comparable number of HCPs over a broad range of isoelectric points and sizes. Importantly, these aptamers detect multiple contaminants that are known to be responsible for therapeutic antibody degradation and toxicity in patients. Because HCP aptamer reagents are sequence-defined and chemically synthesized, we believe they may enable safer production of biologic drugs, and this strategy should be broadly applicable for the generation of HCP detection reagents for other cell lines.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.7b04775