Quantification of amyloid-beta 40 in cerebrospinal fluid.

Autor: Verwey NA; Department of Neurology, VU University Medical Center, Alzheimer Center Amsterdam, P.O. Box 7057, 1007 MB Amsterdam, the Netherlands. n.verwey@vumc.nl, Veerhuis R, Twaalfhoven HA, Wouters D, Hoozemans JJ, Bollen YJ, Killestein J, Bibl M, Wiltfang J, Hack CE, Scheltens P, Blankenstein MA
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2009 Aug 31; Vol. 348 (1-2), pp. 57-66. Date of Electronic Publication: 2009 Jul 01.
DOI: 10.1016/j.jim.2009.06.011
Abstrakt: Background: Truncated forms and full-length forms of the amyloid-beta 40 (Abeta40) are key molecules in the pathogenesis of dementia, and are detectable in CSF. Reliable methods to detect these biomarkers in CSF are of great importance for understanding the disease mechanisms and for diagnostic purposes.
Methods: VU-alpha-Abeta40, a monoclonal antibody (mAb) specifically detecting Abeta40, was generated and characterized by solid and fluid phase ELISA, surface plasmon resonance spectroscopy (SPRS), immunoprecipitation (IP), immunohistochemical and Western blot (WB) analysis. In addition, an ELISA with VU-alpha-Abeta40 as catching and 6E10 as detecting mAbs was set up and validated. This ELISA was used to measure Abeta40 in CSF of controls (N=27), patients with Alzheimer's disease (AD; N=20), frontotemporal lobe dementia (FTLD; N=14), noninflammatory (N=15) and inflammatory (N=15) neurological conditions.
Results: VU-alpha-Abeta40 specifically recognizes Abeta40 with high affinity (K(A)=1.3x10(9) M(-1)) and detects Abeta40 in AD brain specimens. The developed sandwich ELISA has a detection limit of 0.21 ng/mL, a mean recovery of 90%, and an intra- and inter-assay CV of 1.4% and 7.3%. FTLD patients had a lower mean level of Abeta40 (8.8 (1.9) ng/mL) than controls (12.0 (1.7) ng/mL); p<0.01).
Conclusions: VU-alpha-Abeta40 was successfully implemented in an ELISA which enables us to measure Abeta40 accurately in human CSF. Clinical validation revealed lower levels of Abeta40 in FTLD patients. This finding opens new possibilities for early and differential diagnosis of dementia.
Databáze: MEDLINE
Externí odkaz: