DEVELOPPEMENT D'UNE METHODE POUR UTILISER DES NANOPARTICULES DE POLYSTYRENE/DEXTRAN LORS D'UN IMMUNO-ESSAI

DEVELOPMENT OF A METHOD FOR EMPLOYING POLYSTYRENE/DEXTRAN NABO-PARTICLES IN IMMUNOASSAY

Abrégé anglais

A method, for employing Polystyrene/Dextran nanometer particles sized in 5-
300nm in
immunoassay, is established. Polystyrene/Dextran nano-particles are coated
with protein or protein
conjugates which are labeled with fluorescence, enzyme, biotin, magnetic
moterial, luminescence
material or isotope. The nano-particles coated protein or protein conjugates
are used
immunochromatography, immunofiltration and other immunoassay. The nano-
particles of different
sized, coated protein often used analyte, which are arranged like arrays,
coated protein conjugates
used detector which are machine-readable.

Revendications

CLAIMS:
1. A method for immunoassay, which employs nano-meter sized
polystyrene/Dextran
(polystyrene/dextran nano-particle), wherein said polystyrene nano-particles
are coated with
proteins or protein conjugation.
2. The method of claim 1, wherein said polystyrene/Dextran nano-particles are
colored or not
colored.
3. The method of claim 1, wherein said polystyrene/Dextran nano-particles are
divided into at
least one particle group, particles in different groups being coated with
different proteins, for
detecting at least one analyte.
4. The method of claim 1, wherein said polystyrene/Dextran nano-particles have
at least one
size, particles of different sizes being coated with different proteins, for
detecting at least one
analyte.
5. The method of combination of claim 3 and 4.
6. The method of claim 1, 2, 3, 4, 5, wherein said polystyrene / Dextran nano-
particles are in
size range of 5nm to 300nm.
7. The method of claim 1, 2, 3, 4, 5, and 6, wherein said coated proteins are
antigen, antibody,
acceptor, or ligand.
8. An immunoassay, which employs polystyrene/Dextran nano-particle, wherein
said
polystyrene/Dextran nano-particles are coated with proteins.
9. The immunoassay of claim 8, wherein said polystyrene/Dextran nano-particles
are divided
into at least one particle group, particles in different groups being coated
with different
proteins, for detecting at least one analyte.
10. The immunoassay of claim 8, wherein said polystyrene/Dextran nano-
particles have at least
one size, particles of different sizes being coated with different proteins,
for detecting at least
one analyte.
11. The immunoassay of combination of claim 9 and 10.
12. The immunoassay of claim 9, 10 and 11, wherein said polystyrene/Dextran
nano-particles are
colored with at least one color.
13. The immunoassay of claim 8, 9, 11, and 12, wherein said
polystyrene/Dextran nano-particles
are in size range of 5nm to 300nm.

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14. An immunochromatography assay, which employs polystyrene/Dextran nano-
particle,
wherein said polystyrene/Dextran nano-particles are coated with proteins.
15. The immunochromatography assay of claim 14, wherein said
polystyrene/Dextran nano-
particles have at least one size, being divided into at least one particle
group, being in size
range of 5nm to 300nm; said coated proteins are antigen, antibody, acceptor,
or ligand.
16. The immunochromatography assay of claim 15 wherein said assay includes
three areas in
sequence:
a) a sample receiving area, for receiving a liquid sample to be tested;
b) a conjugate area having therein or thereon said polystyrene/Dextran nano-
particles in
dry state;
c) a read-out area having at least one defined zone, wherein on each said
defined zone a
complementary binding partner protein to a protein on said particle is
immobilized.
The immunochromatography assay works by adding the liquid sample to said
sample
receiving area, when said sample passing said conjugate area
polystyrene/Dextran nano-
particles of one or more sizes being released and moving forward together with
said
sample, said sample carrying said released polystyrene/Dextran nano-particles
is then
passing said read-out area, said released polystyrene/Dextran nano-particles
are thus
detected when said sample passes said defined zone(s) in said read-out area,
and the
analyte in the sample are therefore detected qualitatively and/or
quantitatively.
17. The immunochromatography assay of claim 16, wherein said defined zone(s)
at said read-out
area have line shape.
18. The immunochromatography assay of claim 17, wherein said line(s) are
arranged in the
following order:
The complementary binding partner protein(s) immobilized to said line(s)
closer to said
conjugate area correspond to said protein(s) coated on larger size particles
and the closer said
line to said conjugate area the larger said corresponding particle size; on
the contrary, the
complementary binding partner protein(s) immobilized to said line(s) farther
to said
conjugate area corresponds to said protein(s) coated on smaller size particles
and the farther
said line to said conjugate area the smaller said corresponding particle size.
19. The immunochromatography assay of claim 16 wherein said defined zone(s) at
said read-out

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area have spot shape.
20. The immunochromatography assay of claim 16 wherein said defined zones at
said read-out
area have spot shapes, forming a spot matrix.
21. The immunochromatography assay of claim 19 and 20, wherein said spot(s)
are arranged in
the following order:
The complementary binding partner protein(s) immobilized to said spot(s)
closer to said
conjugate area correspond to said protein(s) coated on larger size particles
and the closer said
spot to said conjugate area the larger said corresponding particle size; on
the contrary, the
complementary binding partner protein(s) immobilized to said spot(s) farther
to said
conjugate area corresponds to the protein(s) coated on smaller size particles
and the farther
said spot to said conjugate area the smaller said corresponding particle size.
22. An immunofiltration assay, which employs polystyrene/Dextran nano-
particles, wherein said
polystyrene/Dextran nano-particles are coated with proteins.
23. The immunofiltration assay of claim 22, wherein said polystyrene/Dextran
nano-particles
have at least one size, being divided into at least one particle group, and
being colored with
at least one color, being in size range of 5nm to 300nm; said coated proteins
are antigen,
antibody, acceptor, or ligand.
24. The immunofiltration assay of claim 23, wherein said polystyrene/Dextran
nano-particles are
in aqueous phase, and wherein said assay includes a filter, on which there is
at least one
defined zone on that a complementary binding partner protein to a protein on
said
polystyrene/Dextron nano-particles is immobilized.
The immunofiltration assay works by adding theliquid sample to said filter and
said sample
permeates said filter, said aqueous polystyrene/Dextran nano-particles coated
with proteins
are then added to said filter, said polystyrene/Dextran nano-particles are
thus detected in the
defined zone(s), and the analyte in said sample are therefore detected
qualitatively and/or
quantitatively.
25. The immunofiltration assay of claim 24 wherein said defined zone(s) have
line shape.
26. The immunofiltration assay of claim 24, wherein said defined zone(s) have
spot shape.
27. The immunofiltration assay of claim 24, wherein said defined zones have
spot shapes,
forming a spot matrix.
28. The method of claims 1, 2, 3, 4, 5, 6, and 7 wherein said protein coated
to said

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polystyrene/Dextran nano-particle is labeled with enzyme, fluorescein, biotin,
isotope,
magnetic material, and/or luminescent material.
29. The assay of claims 8, 9, 10, 11, 12, 13, wherein said protein coated to
said
polystyrene/Dextran nano-particle is labeled with enzyme, fluorescein, biotin,
isotope,
magnetic material, and/or luminescent material.
30. The immunochromatography assay of claims 14, 15, 16, 17, 18, 19, 20 and 21
wherein said
protein coated to said polystyrene/Dextran nano-particles is labeled with
enzyme,
fluorescein, bitin, isotope, magnetic material, and/or luminescent material.
31. The immunofiltration assay of claims 22, 23, 24, 25, 26 and 27 wherein
said protein coated
to said polystyrene/Dextran nano-particles is labeled with enzyme;
fluorescein, biotin,
isotope, magnetic material, and/or luminescent material.
32. A method for immunoassay, which employs polystyrene/Dextran nano-particles
in the size of
5nm to 300nm, coated with protein such as antigen or antibody, wherein said
protein is
immobilized to a solid support.
33. A method for immuno-separation, which employs polystyrene/Dextran nano-
particles in the
size of 5nm to 300nm, coated with proteins such as antigen or antibody,
wherein said protein
is labeled by magnetic material.

Description

CA 02489105 2004-12-20
DESCRIPTION:
In immunochromatography assay, Dextran manometer particles is combined with
polystreptavidin,
and they are formed complex. This complex, as capture reagent, is binding on
membrane. The
Biotiny labeled antibody is combined capture reagent, and the test line or
test spot is formed. FITC
Fluorescein labeled antibody coated polystyrene namo-particle, or antibody
coated colored
polystyrene namo-particle or colored latex bead, is as detective reagent.
In immunochromatography assay, when sample go through lateral flow system, if
there are expected
antigen in the sample, Dextran namo-particles polystreptavidin or biotinylated
antibody, and antigen
or colored latex bead are formed complex which are machine-readable or visual
readable. No
antigen in the sample, no complex is formed, and the result is negative.
In immunochromatography assay capture reagent also can be treat antibody
coated namo-particle bind
on membrane. Detective reagent may labeled with fluorescein enzyme, biotin,
isotope, magnetic
material and/or luminesient material. The protein coated namo-particles may be
antibody, antigen,
acceptor or ligend. Polystyrene/Dextran namo-particle sized range are 5-
300nxn.
In immunochromatography assay, the capture reagent binding on membrane may be
single spotlline
or mutil-sports which are arranged matrix sports. In immunofiltration assay,
polystyrene/Dextran
namo-particles coated protein(antibody, antigen, avidin, biotin etc) bind on
membrane as capture
reagent. The antigen or antibody labeled fluorescein, enzyme, magnetic
material or luminesient
material may be coated namo- particle as detective reagent. These namo-
particles sized ranged are 5-
300 mm.
In immunoassay, Dextran namo-particles sized range 5-300 mm, coated with
protein such as antigen,
antibody on streptavidin, combined with protein namo-particle, may be attached
on polystyrene tube
or well, as solid support. The namo-particles in the sized reagent coated
protein which labeled with
flurescein enzyme, magnetic material or luminesient material can be formed a
new solid immune test
model.