An acid dissociation bridging ELISA for detection of antibodies directed against therapeutic proteins in the presence of antigen
Abstract
Bridging Enzyme-Linked Immunosorbent Assay (ELISA) is commonly used in detection of antibodies directed against therapeutic proteins. Advantages of the bridging ELISA include the capability to detect antibodies regardless of their isotype or the species of origin. However, detection of antibodies can be difficult, if not impossible, in the presence of high levels of the antigen in the sample matrix. This protocol describes a bridging ELISA that uses a covalently coupled high density antigen surface combined with an acid dissociation step to allow for antibody detection in the presence of antigen in human serum.
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Currently, the majority of bioanalytical methods used to determine mAb concentrations are ligand-based assays such as ELISA.
“Currently, the majority of bioanalytical methods used to determine mAb concentrations are ligand‐based assays (e.g., ELISA).54”
From Tutorial on monoclonal antibody pharmacokinetics and its considerations in early development (2018) · cites this paper as [54]
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- Authors
- Patton, A., Mullenix, M. C., Swanson, S. J., & Koren, E.
- Affiliations
- Amgen (United States) (US)
- Corresponding author
- Koren, Eugen
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 2005-09
- Volume
- 304
- Issue
- 1-2
- Pages
- 189-195
- ISSN
- 0022-1759 (ISSN-L)
- OpenAlex
- W1997496599
- Cited by
- 201
- References
- 17
- Citations by year
- 2026: 6 · 2025: 8 · 2024: 12 · 2023: 5 · 2022: 13 · 2021: 3 · 2020: 8 · 2019: 5 · 2018: 9 · 2017: 12 · 2016: 15 · 2015: 12 · 2014: 15 · 2013: 15 · 2012: 11
- FWCI
- 4.29
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Immunological Methods (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Immunology and Microbiology › Immunology › Biosimilars and Bioanalytical Methods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Keywords
- anti-drug antibodies, therapeutic proteins, acid dissociation
- MeSH terms
- Antibodies, Antigens, Calibration, Enzyme-Linked Immunosorbent Assay, Humans, Proteins
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