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Generation of mid‐region murine monoclonal antibodies against multiple structural isoforms of amyloid‐beta by hybridoma technology
Author(s) -
Stockmann Julia,
Beyer Léon,
Galkowski Sandy,
Woitzik Nathalie,
Güldenhaupt Jörn,
Gerwert Klaus Bernhard
Publication year - 2020
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1002/alz.043577
Subject(s) - polyclonal antibodies , antibody , monoclonal antibody , microbiology and biotechnology , chemistry , biomarker , epitope , peptide , antigen , immunology , medicine , biology , biochemistry
Abstract Background The immuno‐infrared‐sensor enables the diagnosis of Alzheimer’s disease by measuring the secondary structure distribution of Amyloid‐beta (Aß) in CSF and blood plasma. For this technique an antibody is essential, which binds to all conformations of the biomarker peptide. We developed different immunization strategies to generate monoclonal Aß‐antibodies (mAb) using wt and APP Knock‐Out (KO) mice. Method KO and wt mice were immunized with different Aß peptides. With an antigen‐specific ELISA antibody‐producing hybridoma cells were screened and selected for performance tests with the immuno‐infrared sensor. Immuno‐infrared‐analyses provided a detailed characterization of the structural selectivity of the antibodies. Result Both KO and wt mice generated multiple Aß specific antibodies, though KO mice yielded 8 times more Aß‐positive antibodies than wt. All antibodies, probably polyclonal, revealed binding to alpha‐helical or disordered monomeric Aß as well as to ß‐sheet enriched Aß (oligomers, fibrils). Additionally, seven of these antibodies differentiated between AD patients and healthy controls in cerebrospinal fluid (CSF) samples. Conclusion With our immunization strategy we were able to generate multiple antibodies for the diagnosis of Alzheimer’s disease with the immuno‐infrared sensor. We generated seven antibodies, which enable the differentiation of AD patients and healthy controls by analyses on CSF. References: [1] Nabers, A., Ollesch J. et al. (2016) J. Biophotonics, 9. 224‐234 . [2] Nabers, A. et al. (2018) EMBO Mol. Med. 10, e8763 . [3] Nabers, A et al. (2019) Alzheimer’s Dement. Diagnosis, assess. Dis. Monit., 11, 257‐263 . [4] Stocker, H et al. (2019), Alzheimer's & Dementia , in press.