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1. WO2020115356 - AMPLIFICATION PAR TRANSPORT À MÉDIATION PAR L'AFFINITÉ

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS

1. A method for amplification in a microfluidic assay for analyte detection,

comprising

binding an analyte (90) to a transport element, wherein the transport element comprises a particle (70) and a first antibody element (80) and the analyte is bound to the first antibody element (80);

moving the transport element with the analyte (90) bound thereto towards a tracer storage site (10b) comprising tracer elements, wherein a tracer element comprises a first binder element (30), a label element (40) and a second antibody element (50);

binding the analyte to the second antibody element (50);

moving the transport element with the analyte (90) and the tracer element bound therewith towards a tracer transport site (10a) comprising second binder elements (20);

binding the first binder element (30) with a second binder element (20); moving the transport element with the analyte (90) bound thereto towards the tracer storage site (10b) so that the analyte is detached from the second antibody element (50) of the tracer element while the first binder element (30) remains bound to the second binder element (20); and

repeating the steps subsequent to binding the analyte (90) to the transport element a predetermined number of times; wherein

the first antibody element (80) has a higher affinity for the analyte (90) than the second antibody element (50);

the second antibody element (50) has a higher affinity for the analyte (90) than for the tracer storage site (10b); and

the affinity of the second antibody element (50) towards the analyte (90) is smaller than that between the first (30) and second (20) binder elements and that between the second antibody element (50) and the first binder element (30).

2. The method of claim 1 , wherein moving the transport element comprises causing a force that moves the transport element.

3. The method of claim 1 or 2, wherein moving the transport element comprises causing a magnetic force and the particles (70) of the transport element comprise magnetic nanoparticles (70).

4. The method of any preceding claim, wherein the first binder element (30)

comprises biotin and the second binder element (20) comprises avidin.

5. The method of any preceding claim, wherein the label element (40) comprises a fluorescent dye.

6. A method for detecting an analyte in a microfluidic assay, comprising

introducing a fluid comprising the analyte (90) to be detected into the assay;

carrying out the amplification method of any preceding claims; and subsequently

reading out the result of the assay.

7. The method of claim 6, wherein reading out the result of the assay comprises fluorescent excitation and optical detection of the resulting fluorescence.

8. An apparatus for a microfluidic assay for analyte detection, comprising

a microfluidic chip (410), comprising at least one input port (110) for introducing a fluid containing an analyte (90), and at least one assay site (100), the assay site comprising transport elements, a tracer transport site (10a) and a tracer storage site (10b); wherein

a transport element comprises a particle (70) and a first antibody element (80) and the analyte is bound to the first antibody element (80);

the tracer storage site (10b) comprises tracer elements comprising a second antibody element (50) bound to a first binder element (30) and a label element (40);

the tracer transport site (10a) comprises second binder elements (20) for binding with a first binder element (30); characterized in that

the transport element is configured to be movable towards the tracer transport site (10a) and the tracer storage site (10b); and in that

the first antibody element (80) has a higher affinity for the analyte (90) than the second antibody element (50);

the second antibody element (50) has a higher affinity for the analyte (90) than for the tracer storage site (10b); and

the affinity of the second antibody element (50) towards the analyte (90) is smaller than that between the first (30) and second (20) binder elements and that between the second antibody element (50) and the first binder element (30).

9. The apparatus of claim 8, wherein the particle (70) comprises a magnetic

nanoparticle.

10. A system for a microfluidic assay for analyte detection, comprising

the apparatus of any of the claims 8-9; and

a processor configured to cause carrying out the method of any of the claims 1 -7.

11. The system of claim 10 further comprising a control element (410) configured to provide the force for moving the transport elements and to handle the fluids in the assay.

12. The system of claim 10 or 11 , further comprising a detection element (430)

configured to read out the result of the assay.