Generation and Partial Characterization of Rabbit Monoclonal Antibody to Amyloid-β Peptide 1–37 (Aβ₃₇)

Article type: Research Article

Authors: Mehta, Pankaj D.^{a; *} | Blain, Jean-Francois^b | Freeman, Emily A.^b | Patrick, Bruce A.^a | Barshatzky, Marc^a | Hrdlicka, Lori A.^b | Mehta, Sangita P.^a | Frackowiak, Janusz^a | Mazur-Kolecka, Bozena^a | Wegiel, Jerzy^a | Patzke, Holger^b | Miller, David L.^a

Affiliations: [a] New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York, NY, USA | [b] FORUM Pharmaceuticals, Waltham, MA, USA

Correspondence: [*] Correspondence to: Pankaj D. Mehta, PhD, Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, NY 10314-6315, USA. Tel.: +1 718 494 5159; Fax: +1 718 982 634; E-mail: [email protected].

Abstract: Secreted soluble amyloid-β 1–37 (Aβ37) peptide is one of the prominent Aβ forms next to Aβ40, and is found in cerebrospinal fluid (CSF) and blood. Recent studies have shown the importance of quantitation of CSF Aβ37 levels in combination with Aβ38, Aβ40, and Aβ42 to support the diagnosis of patients with probable Alzheimer’s disease (AD), and the value of antibody to Aβ37 to facilitate drug discovery studies. However, the availability of reliable and specific monoclonal antibody to Aβ37 is very limited. Our aims were: 1) to generate and partially characterize rabbit monoclonal antibody (RabmAb) to Aβ37, and 2) to determine whether the antibody detects changes in Aβ37 levels produced by a γ-secretase modulator (GSM). Our generated RabmAb to Aβ37 was found to be specific to Aβ37, since it did not react with Aβ36, Aβ38, Aβ39, Aβ40, and Aβ42 in an ELISA or immunoblotting. The epitope of the antibody was contained in the seven C-terminal residues of Aβ37. The antibody was sensitive enough to measure CSF and plasma Aβ37 levels in ELISA. Immunohistological studies showed the presence of Aβ37-positive deposits in the brain of AD, and Down syndrome persons diagnosed with AD. Our studies also showed that the antibody detected Aβ37 increases in CSF and brains of rodents following treatment with a GSM. Thus, our antibody can be widely applied to AD research, and in a panel based approach it may have potential to support the diagnosis of probable AD, and in testing the effect of GSMs to target AD.

Keywords: Alzheimer disease, amyloid-β peptide, Down syndrome, ELISA, immunoblotting, immunohistochemistry, rabbit monoclonal antibodies to Aβ₃₇

DOI: 10.3233/JAD-161207

Journal: Journal of Alzheimer's Disease, vol. 57, no. 1, pp. 135-145, 2017