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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| Published in | Journal of bioscience and bioengineering Vol. 126; no. 4; pp. 522 - 526 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
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Japan
Elsevier B.V
01.10.2018
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| Abstract | 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.
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| AbstractList | 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] 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. |
| Author | Muneoka, Satoshi Utsugisawa, Kimiaki Makino, Tomohiro Nakamura, Ryuichi Hoshino, Masato |
| Author_xml | – sequence: 1 givenname: Satoshi orcidid: 0000-0001-6326-0983 surname: Muneoka fullname: Muneoka, Satoshi email: muneoka.satoshi.j7@daiichisankyo.co.jp organization: Asubio Pharma Co., Ltd., 6-4-3 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan – sequence: 2 givenname: Ryuichi surname: Nakamura fullname: Nakamura, Ryuichi organization: Asubio Pharma Co., Ltd., 6-4-3 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan – sequence: 3 givenname: Masato surname: Hoshino fullname: Hoshino, Masato organization: Asubio Pharma Co., Ltd., 6-4-3 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan – sequence: 4 givenname: Kimiaki surname: Utsugisawa fullname: Utsugisawa, Kimiaki organization: Department of Neurology, Hanamaki General Hospital, 4-28 Kajoh-chou, Hanamaki, Iwate 025-0075, Japan – sequence: 5 givenname: Tomohiro surname: Makino fullname: Makino, Tomohiro organization: Asubio Pharma Co., Ltd., 6-4-3 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/29857948$$D View this record in MEDLINE/PubMed |
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| Keywords | AlphaLISA Myasthenia gravis Antibody screening Nicotinic acetylcholine receptor Membrane proteins |
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| SubjectTerms | 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 |
| Title | Development of a novel immunoassay to select antibodies against intact membrane antigens by using the homogeneous AlphaLISA system |
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