Polymer microparticle-based method for probe deposition in label-free biosensors
Disclosed is a sensor chip for detecting a target molecule in a sample. The sensor chip includes a substrate having a surface and a layer of hydrogel particles immobilized on the substrate surface at two or more locations on the surface, wherein the hydrogel particles at a first location comprise a plurality of first probe molecules bound to the particles and the hydrogel particles at a second location comprise a plurality of second probe molecules bound to the particles. Systems that include the sensor chip, as well as methods of preparing and using the sensor chip, are also disclosed.
1. A sensor chip for detecting a target molecule in a sample, the sensor chip comprising:
a substrate comprising a coating compatible with label-free, optical detection of probe molecule-target molecule binding, the substrate and coating each having a refractive index and the coating having a surface and a thickness; and
a monolayer of hydrogel particles immobilized on the coating surface at two or more locations on the surface, wherein the hydrogel particles at a first location comprise a plurality of first probe molecules bound to the particles and the hydrogel particles at a second location comprise a plurality of second probe molecules bound to the particles;
wherein the sensor chip is an Arrayed Imaging Reflectometry sensor chip, the substrate is a silicon substrate, and the coating is a silicon dioxide coating having a surface roughness of less than about 0.5 nm across the substrate surface, and
wherein, due to the combination of the silicon substrate and silicon dioxide coating refractive indices and silicon dioxide coating thickness, the structure of the sensor chip affords, upon illumination of the sensor chip with s-polarized light having a particular wavelength and a particular angle of incidence, a near zero background reflectivity in the absence of probe molecule-target molecule binding at any of the two or more locations and increased reflectivity in the presence of probe molecule-target molecule binding at any of the two or more locations.
2. The sensor chip according to claim 1 , wherein the hydrogel particles are non-covalently attached to the substrate surface.
3. The sensor chip according to claim 1 , wherein the hydrogel particles comprise poly-N-isopropylacrylimide (PNIPAM), PNIPAM copolymerized with allyl-iminodiacetic acid, PNIPAM copolymerized with vinylacetic acid, PNIPAM copolymerized with allylamine, PNIPAM grafted with polyethylene glycol-succinic acid, hydroxypropyl cellulose, or a pullulan acetate/sulfonamide conjugate.
4. The sensor chip according to claim 1 , wherein one or both of the first and second probe molecules are covalently bound to the hydrogel particles of first and second locations, respectively.
5. The sensor chip according to claim 1 , wherein one or both of the first and second probe molecules are non-covalently bound to the hydrogel particles of first and second locations, respectively.
6. The sensor chip according to claim 1 , wherein the monolayer of hydrogel particles at the two or more locations is characterized by one or more of:
particles having an average diameter of about 50 nm to about 1000 nm;
particle densities of about 10 8 to about 10 14 particles/mL per volume applied to each location;
a monolayer thickness of less than 10 nm; and
a density of first and/or second probe molecules of about 1 to about 10 6 per particle.
7. The sensor chip according to claim 1 , wherein the first and second probe molecules are independently selected from the group consisting of oligonucleotides, nucleic acid aptamers, peptides, proteins, and non-polymeric small molecules.
8. The sensor chip according to claim 1 , wherein the sensor chip is capable of detecting the probe molecule-target molecule binding while the monolayer of hydrogel particles remain immobilized on the substrate surface.
9. A detection system comprising:
a sensor chip according to claim 1 ;
a light source that is positioned to illuminate the sensor chip with s-polarized light; and
a detector that is positioned to detect light reflected or emitted from the surface of the sensor chip, and thereby determine whether a target molecule is present in a sample exposed to the surface of the sensor chip.
10. A method for sensing a target molecule in a sample, the method comprising:
providing a detection system according to claim 9 ;
directing s-polarized light at a surface of the sensor chip;
contacting the sensor chip with a sample under conditions effective to allow a target molecule in the sample to bind specifically to one of the first and second probe molecules; and
detecting light reflected or emitted by sensor chip to identify the presence of a target molecule in the sample.
11. The method according to claim 10 , wherein the sensor chip of the provided detection system is dry and the method further comprises wetting the sensor chip prior to said contacting.