Arrays, substrates, devices, methods and systems for detecting target molecules
Arrays and substrates comprising a material, in particular capture agents and/or detectable targets, attached to the substrates along substantially parallel lines forming a barcoded pattern and related methods and systems.
1. A method for multiplexed sample analysis, the method comprising:
providing a device comprising:
a surface retaining at least four separate lines of capture antibodies arranged parallel to one another and in a serpentine-like pattern over the surface, wherein each line of capture antibodies comprises a specific and different antibody, relative to antibodies of adjacent lines of capture antibodies, configured to bind to a different target molecule;
and
a substrate that couples with the surface, the substrate comprising a plurality of open chambers, wherein:
the coupling of the substrate and the surface is such that a portion of the surface covers and encloses each chamber of the plurality of chambers to form a plurality of respective enclosures resulting in each enclosure of the plurality of enclosures being enclosed on all sides,
each enclosure of the plurality of enclosures includes a biological sample,
the biological sample comprises one or more components selected from the group consisting of a biological fluid, a cell, a plurality of cells, a tissue, and a combination thereof,
the portion of the surface covering each chamber of the plurality of chambers to form each enclosure of the plurality of enclosures includes at least one antibody set comprising a portion of each of the lines of capture antibodies attached to the surface;
providing reaction conditions that allow for specific interaction between the one or more components of the biological sample and at least one capture antibody from the plurality of lines of capture antibodies, or, between at least one target molecule of the one or more components of the biological sample and the at least one capture antibody, so as to form at least one capture antibody complex;
providing at least one labeled secondary antibody that specifically binds to the at least one capture antibody complex, wherein:
the at least one labeled secondary antibody detects a specific interaction between the at least one capture antibody and the one or more components of the biological sample or between at least one target molecule and the at least one capture antibody, providing a detectable signal corresponding to the specific interaction, and
the detectable signal comprises a signature corresponding to the biological sample; and
visualizing at least one signature to identify one or more components of the biological sample.
2. The method of claim 1 , wherein at least one target molecule is present in at least a portion of the plurality of enclosures.
3. The method of claim 1 , wherein at least one target molecule is present in each enclosure of the plurality of enclosures.
4. The method of claim 1 , wherein the one or more components of the biological sample includes one or more cell(s) and at least one target molecule is produced by the one or more cell(s).
5. The method of claim 4 , wherein at least one target molecule comprises a chemokine, a cytokine, a growth factor, or a transcription factor.
6. The method of claim 1 , wherein at least one target molecule comprises a chemokine, a cytokine, a growth factor, or a transcription factor.
7. The method of claim 1 , wherein the biological sample includes at least one target molecule and the biological sample is provided in a buffer solution.
8. The method of claim 1 , wherein the biological sample includes at least one target molecule and a biological fluid.
9. The method of claim 8 , wherein the biological fluid comprises blood, plasma, serum, saliva, joint fluid, cell lysate, or tissue lysate.
10. The method of claim 1 , wherein the biological sample includes a component of at least one cell.
11. The method of claim 1 , wherein the labeled secondary antibodies comprise fluorescent, gold or silver labels.
12. The method of claim 1 , wherein the method further comprises quantifying one or more components of the at least one signature.
13. The method of claim 12 , wherein the quantifying step comprises measuring an intensity and/or a density of the labeled secondary antibody.
14. The method of claim 1 , wherein the separate lines of capture antibodies comprises between 4 and 50 separate lines of capture antibodies, each of which specifically bind to a different target molecule.
15. The method of claim 1 , wherein the at least one signature indicates a health or disease state of a subject.
16. The method of claim 1 , wherein the method further comprises analyzing two or more signatures to generate a pattern of expression and/or abundance of one or more components of the biological sample.