Method for making protein chip by combination of magnetic beads and quantum dots
A protein chip, quantum dot technology, applied in measurement devices, material testing products, instruments, etc., can solve problems such as difficulty in maintaining protein activity, being easily affected by liquid environment, and inconvenient for regeneration and reuse.
Inactive Publication Date: 2015-07-01
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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Problems solved by technology
The physical adsorption method has wide applicability, generally adopts spotting technology, the array density is small, the binding between the spot and the substrate is not firm, and it is easily affected by the liquid environment, and the protein is easily inactivated
The chemical cross-linking method is firm, but it has poor versatility for ever-changing proteins, and it is not easy to regenerate and reuse, and it is also difficult to maintain protein activity.
Affinity binding immobilization is of course better in firmness, and the accuracy of the constructed chip is the best, but it has the worst applicability because it needs the specific binding of the stationary phase and the substance to be tested.
[0006] Traditional protein chip detection methods are increasingly unable to meet the technical requirements of complex biochips. The use of fluorescent detection of protein chips labeled with quantum dots modified with thioglycolic acid provides a fast, reliable and cheap detection method for biochip detection.
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Embodiment Construction
[0013] The first step is to prepare complexes of magnetic beads, quantum dots and proteins.
[0014] Select quantum dots and magnetic beads whose surface chemical groups are complementary in reaction, and carry out a chemical reaction to make them cross-linked together through chemical bonds to form a magnetic bead-quantum dot complex. During the reaction, the quantum dots were added in excess to ensure that the magnetic beads were all bound to the quantum dots.
[0015] A magnet is used to separate the cross-linked magnetic bead-quantum dot complex from the solution, and to wash and purify.
[0016] Mix the magnetic bead-quantum dot complex with the protein solution to make it chemically react. During the reaction, the protein is added in excess to ensure that the magnetic bead-quantum dot complex is fully bound to the protein.
[0017] The ternary complex of magnetic beads, quantum dots and protein is separated from the solution by using a magnet, washed and purified for u...
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Abstract
The invention discloses a method for making a protein chip by combination of magnetic beads and quantum dots. The method comprises that proteins are immobilized and arrayed by coalitions of magnetic beads and quantum dots to form the protein chip. The method solves the problem that the traditional method has a small application range, produces protein activity loss, is inconvenient for detection and cannot realize chip regeneration and recycle. The method comprises the following steps of immobilizing a paramagnetic substance in pits of a chip substrate with a pit array, simultaneously bonding magnetic beads, quantum dots and proteins by a chemical method to obtain a complex, feeding the protein complex through a loose gel layer under the action of a magnetic field, and finally immobilizing the protein complex in the pits of the substrate so that outward oriented immobilization of the chip proteins is realized. A protein buffer can also be arranged in the pit and is used for preventing protein inactivation.
Description
Technical field: [0001] The invention relates to a method for making a protein chip by using a combination of quantum dots and magnetic beads to immobilize and array proteins. Background technique: [0002] After the birth of micron or nanoscale magnetic beads and quantum dots, they quickly became sensitive and efficient tools for the detection or preparation of proteins, nucleic acids, genes, organelles, bacteria or viruses in the field of biological research. [0003] Some magnetic substances have superparamagnetism after being made into nanoparticles, which can be strongly attracted by a magnetic field, but the nanoparticles are not magnetic after the magnetic field is removed. Based on the characteristics of nano-magnetic beads, the surface of the magnetic beads is coated with some organic or inorganic substances to make the surface have specific chemical groups, such as amino, hydroxyl, carboxyl, sulfhydryl, etc. After adding them to protein and other solutions, they ca...
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IPC IPC(8): G01N33/68
CPCG01N33/6803
Inventor 仓怀兴张力段秉亚马建华
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES