NOVEL ELISA TESTING SYSTEM

NOUVEAU SYSTEME D'ESSAI DU TYPE ELISA

English Abstract

Enzyme-linked immunosorbent assay (ELISA) is perhaps one of the most
widespread
assay methods used extensively in the field of immunology. This assay works on
the
basis of known amounts of antigens or antibodies adhered onto a solid surface
made of
polystyrene. The polystyrene-made wells are opened at only one side and each
well is
eight mm in diameter, ten mm in depth and holds up to a maximum of 300 µL,
in volume.
Every 8 or 12 wells joins together to form a 96-well or a 48-well combination
unit. The
inner wall of each well is smooth with a transparent flat base. Generally,
approximately
100 µL of known antigens or antibodies is added to each well occupying one-
third of the
well.

Known amounts of antigens or antibodies are allowed to adhere onto the
polystyrene
surface at a specific temperature for over 10 hours. This process is often
known as
"coating". Following this, protein solutions at high concentrations or
solutions with
concentrated proteins and glucose are allowed to further adhere onto the
polystyrene
wells to ensure maximum occupancy of the well surfaces. In addition, this
second
coating offers better protection on the activity of the antigens or
antibodies. This second
process is termed "blocking".

The commonly used enzyme (horseradish peroxidase) can be linked either to the
antigens
or the antibodies, but in either case the solid-phase reactant is unlabeled.
ELISA can be
made quantitative by setting it up as a competitive inhibition assay in which
free or
unknown amounts of antigens (or antibodies) in samples compete with enzyme-
labeled
antigens (or antibodies) for binding sites on the polystyrene solid phase
coated with
known amounts of antigens or antibodies. Following appropriate washings, the
concentration of enzyme bound to the immune complex is estimated by the
addition of its
substrate that changes colour when the substrate is acted on by the enzyme.
This highly-
specific immunoassay has thusfar become one of the most popular immunoassays
in
immunology and immunology-related studies.

Claims

CLAIMS:

The embodiments of the invention in which an exclusive property or privilege
is claimed
are defined as follows:

a. The creation of grooves to increase the surface area of all polystyrene
adhering
surfaces;

b. The entire reaction process of this new ELISA system is carried out under a
shaking
condition at a specific frequency;

c. This new ELISA system adopts a liquid-rotation washing method on all
washable
polystyrene surfaces.

Description

CA 02300687 2000-02-23
SPECIFICATION:
This patent application relates to the development of a novel ELISA operating
system in
contrast to the operation of a conventional ELISA system. This innovative idea
arises
during the process of investigation and research on the use of an immuno-solid
dry
reagents system with the fully automatic analytical machine.
To fully optimize the adherence capacity of the antigens or antibodies, the
surface area is
increased by creating grooves throughout the lower half inner walls of the
polystyrene
wells. This new ELISA system uses fully solidified reagents and has two
distinct
differences from the conventional ELISA system in terms of operation. Firstly,
the
reactions of both the unknown number of antigens (or antibodies) in samples
and the
enzyme-labeled antigens (or antibodies) are stationary using the conventional
ELISA
method. In this new ELISA system, however, the reactions of both the samples
and
enzyme-labeled antigens (or antibodies) are carried out under shaking at a
specific
temperature. In addition, the shaking frequency varies with different cases or
examples.
Secondly, the washing method following the reactions of both the samples and
enzyme-
labeled antigens (or antibodies) is also different. The conventional ELISA
system adopts
a vertical flow washing mode whereas this new ELISA system uses a liquid-
rotation
washing method.
This patent application pertains to the use of immuno-solid dry reagents
system in
conjunction with the fully automatic analytical machine. The novel idea of a
special
glass-frosting treatment and unique washing method are both creative changes
in this
new ELISA testing system. These unique treatment and changes will definitely
increase
the overall sensitivity and specificity of a more advanced ELISA system.