TITLE:
Combination of specific monoclonal antibodies allow identification of soluble aggregates of by sandwich ELISA
AUTHORS:
Takenori Shimizu, Kazuaki Yoshimune, Tomoe Komoriya, Takahiro Akiyama, Xujun Ye, Hideki Kohno
KEYWORDS:
Aβ1-42; Monoclonal Antibody; Soluble Aggregates; ELISA
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.4 No.4A,
April
30,
2013
ABSTRACT:
Aggregate amyloid beta protein1-42 (Aβ1-42) can typically be
found in the early stage of Alzheimer’s disease (AD). Aβ1-42 self-assembles and is highly toxic to neurons.
Thus, recognizing aggregated Aβ1-42 is very important for elucidation of Aβ1-42 structure and for the diagnosis of AD. In this study, the specificity of the
79-3 monoclonal antibody against soluble aggre- gate Aβ1-42 was measured by sandwich Enzyme-Linked Immuno Sorbent
Assay (ELISA). Eight monoclonal antibodies against both soluble aggregates
and amorphous aggregates were used as primary antibodies. Soluble aggregates
and amorphous aggregates were used as antigen. As secondary antibody, HRP was
labeled with the 79-3 monoclonal antibody. The reactivity of the 79-3
monoclonal antibody against soluble aggregates was confirmed in all combinations,
but little reactivity against amorphous aggregates was found. Furthermore, we
performed the above sandwich ELISA using the 37-11 antibody, which is reactive
against large oval aggregates (LOA) that
occur in micro aggregates, instead of the 79-3 antibody. The 77-3 antibody is 1 of the 8 monoclonal antibodies against soluble aggregates; amorphous aggregates also reacted with the
37-11 antibody. These results indicated that soluble aggregates are specifically
recognized by a combination of different antibodies. The combined use of these
antibodies can be applied to the diagnosis of AD and to defining the structure
of the Aβ1-42.