Rapid Quantitation of Various Therapeutic Monoclonal Antibodies Using Membranes with Fc-Specific Ligands

Therapeutic monoclonal antibodies (mAbs) provide effective treatments for many diseases, including cancer, autoimmune disorders, and, lately, COVID-19. Monitoring the concentrations of mAbs is important during their production and subsequent processing. This work demonstrates a 5 min quantitation of...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2023-06, Vol.95 (22), p.8541-8551
Main Authors: Yang, Junyan, Ostafe, Raluca, Welch, Christopher J., Verhalen, Brandy, Budyak, Ivan L., Bruening, Merlin L.
Format: Article
Language:English
Subjects:
Acceptance criteria
Antibodies, Monoclonal
Assaying
Autoimmune diseases
Binding
Broths
Carboxylic acids
Chemistry
COVID-19
Enzyme-Linked Immunosorbent Assay - methods
Fermentation
Fluorescence
Humans
IgG antibody
Immunoassays
Immunoglobulin G
Ligands
Lysates
Membrane proteins
Membranes
Monitoring
Monoclonal antibodies
Polyelectrolytes
Protein A
Proteins
Quantitation
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Summary:Therapeutic monoclonal antibodies (mAbs) provide effective treatments for many diseases, including cancer, autoimmune disorders, and, lately, COVID-19. Monitoring the concentrations of mAbs is important during their production and subsequent processing. This work demonstrates a 5 min quantitation of most human immunoglobulin G (IgG) antibodies through capture of mAbs in membranes modified with ligands that bind to the fragment crystallizable (Fc) region. This enables binding and quantitation of most IgG mAbs. Layer-by-layer (LBL) adsorption of carboxylic acid-rich polyelectrolytes in glass-fiber membranes in 96-well plates allows functionalization of the membranes with Protein A or a peptide, oxidized Fc20 (oFc20), with high affinity for the Fc region of human IgG. mAb capture occurs in
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.3c00531