LUCIO®-Medical ELISA Brochure - nal von minden

Test Principle
Our LUCIO®-Medical and Sero ELISA are all based
on the principle of antibody-antigen binding,
which results in an extremely high sensitivity.
Thereby, specially developed monoclonal antibodies
(MAb), which specifically recognize an
epitope of an antigen, are used causing a very
high specificity.
To be able to quantitatively evaluate the extent
of the antibody-antigen binding, an enzyme
(mostly horseradish-peroxidase) changing a
colorless substrate into a colored product, is
used. Depending on the issue in question or on
the molecule of interest, this principle is performed
in different ways.
Antibody-Enzyme-
Conjugate
Antigen
from
the
sample
Antigen
from
the
sample
Antigen-Enzyme-
Conjugate
1. Direct Detection of the Antigen
Antibodies against the antigen of interest are
bound to the microtiter plate. The binding of
the antigen is visualized either by means of
an antibody-enzyme conjugate (Sandwich-
ELISA) or by a competing antigen-enzyme
conjugate (competitive ELISA).
Sandwich ELISA for
antigen detection
Competitive ELISA for
antigen detection
Anti-IgG-Antibody-
Enzyme-Conjugate
IgG-Antibody
from the
sample
Sandwich ELISA for the
detection of IgG-Antibodies
Anti-IgM/IgA-
Antibody-Enzyme-
Conjugate
IgM-Antibody
from the
sample
Sandwich ELISA for the
detection of IgM/IgA-Antibodies
2. Direct Detection of the Antibody
Antigens are bound to the microtiter plate.
Antibodies of interest from the sample (IgG,
IgM or IgA) bind to this antigen. The binding
is visualized by using an antibody-enzymeconjugate
that recognizes all bound human
antibodies. Since IgG-antibodies are present
in abundance, they would superimpose the
detection of IgM and IgA antibodies - for this
reason, an ELISA sorbent, which removes all
IgG antibodies from the sample, has to be
used to detect IgM and IgA antibodies.
Antigen-Enzyme-
Conjugate
IgM-Antibody
from the sample
Anti-IgM-
Antibody
3. Indirect Detection of IgM-Antibodies
(µ-capture)
For the indirect detection of IgM-antibodies
by means of a µ-capture, firstly all IgMantibodies
in the samples are bound to
anti-IgM antibodies on the microtiter plate.
The specific detection of the IgM-antibodies
of interest is made by the binding of an
enzyme-linked antigen. This method does
not require the use of a sorbent.
µ-capture ELISA
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