Development of a novel immunoassay to select antibodies against intact membrane antigens by using the homogeneous AlphaLISA system

Membrane proteins, such as G-protein-coupled receptors and ion channels are attractive targets for antibody-based therapeutics as pharmaceutical and biotech companies have increasingly moved their attention to biologics. However, lack of appropriate screening systems to correctly detect specific ant...

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Bibliographic Details
Published in:Journal of bioscience and bioengineering 2018-10, Vol.126 (4), p.522-526
Main Authors: Muneoka, Satoshi, Nakamura, Ryuichi, Hoshino, Masato, Utsugisawa, Kimiaki, Makino, Tomohiro
Format: Article
Language:English
Subjects:
AlphaLISA
Antibodies - analysis
Antibodies - immunology
Antibody screening
Bungarotoxins - analysis
Bungarotoxins - immunology
Humans
Immunoassay - instrumentation
Immunoassay - methods
Membrane proteins
Myasthenia gravis
Nicotinic acetylcholine receptor
Receptors, Nicotinic - analysis
Receptors, Nicotinic - immunology
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Summary:Membrane proteins, such as G-protein-coupled receptors and ion channels are attractive targets for antibody-based therapeutics as pharmaceutical and biotech companies have increasingly moved their attention to biologics. However, lack of appropriate screening systems to correctly detect specific antibodies against membrane proteins has hampered antibody discovery and development so far. In the present study, we described the development of a novel high-throughput immunoassay platform based on AlphaLISA to screen antibodies against intact membrane proteins, taking nicotinic acetylcholine receptor (nAChR), one of the best-known ion channel membrane proteins, as an example. By using signal transfer between α-bungarotoxin, the ligand of the receptor, conjugated with donor beads, and anti-nAChR antibodies (mAb35 and mAb210) with acceptor beads, we could detect strong and specific signals, directly from the homogenates of cells expressing nAChR. Using this platform, we isolated a new human IgG antibody against nAChR in a high-throughput manner. This methodology can be applied for the discovery of antibodies against other types of membrane proteins. [Display omitted]
ISSN:1389-1723
1347-4421
DOI:10.1016/j.jbiosc.2018.04.018