Biomolecule fixing method

A technology of biomolecules and immobilization methods, applied in chemical instruments and methods, semi-permeable membrane separation, membrane technology, etc. Problems such as poor performance, to achieve stable fixation, high yield and good reproducibility

Inactive Publication Date: 2012-11-28
NANJING NORMAL UNIVERSITY
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  • Description
  • Claims
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Problems solved by technology

However, the adsorption method is unstable, the reproducibility of the combination method is poor, the operation process of the covalent bonding method is relatively complicated, and the amount of glutaraldehyde in the cross-linking method is difficult to control. The form exists on the surface of the electrode, so it is difficult for the biomolecules to form a monolayer orderly arrangement on the modified electrode.

Method used

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Embodiment 1

[0025] Embodiment 1 The immobilization method of horseradish peroxidase (HRP)

[0026] (1) Preparation of nanoscale gold

[0027] AuCl 3 .HCl.4H 2 O (2.49 mg, 6.05 mmoL) was mixed with triphenylphosphine (1.6 mg, 6.05 mmoL), and after mixing, toluene (5 mL) was added to disperse and dissolve under ultrasound to obtain organic phase A; CTAB (0.8 mg, 2.2 mmoL) was added to 8 mL Double distilled water, after dissolving, add NaOH (2 mg, 50 mmoL) to completely dissolve the aqueous phase B; slowly pour the organic phase A into the aqueous phase B, after it is stable, add the reducing agent hydrazine hydrate to the aqueous phase B (25 mg, 515 mmoL ), the reaction occurred immediately, and it was o After placed under C condition for 24 hours, the interface has obvious purple-black material generation, that is, nano-gold, such as figure 1 shown.

[0028] (2), preparation of gold and HRP composite film

[0029] HRP (1.5 mg) was dissolved in 1mL PBS (pH 7.0), and then added to the ...

Embodiment 2

[0032] The immobilization method of embodiment 2 DNA

[0033] (1) Preparation of nano-scale silver

[0034] will be dissolved with Ag 2 (PPh 3 ) 4 Cl 2The toluene solution is used as the organic phase, and the alkaline solution is used as the water phase. Add the organic phase to the water phase. After the two phases are stable, inject the tetrahydroxymethylphosphorus chloride (THPC) solution into the water layer with a syringe and let it stand for a while. , nano-silver can be obtained on the liquid-liquid interface. For specific methods, see literature (C.N.R. Rao, G.U. Kulkarni, Ved Varun Agrawal, Ujjal K. Gautam, Moumita Ghosh, Usha Tumkurkar. Use of the liquid–liquid interface for generating ultrathin nanocrystalline films of metals, chalcogenides, and oxides. Journal of Colloid and Interface Science 289 (2005) 305–318).

[0035] (2), preparation of silver and DNA composite film

[0036] DNA was dissolved in PBS, and then added to the prepared nano-silver aqueous p...

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Abstract

The invention provides a novel simple biomolecule fixing method, comprising the following steps: firstly synthesizing a metal nano material by a liquid-liquid interface method; then adding biomolecules to a water phase for fixing the biomolecules to the liquid-liquid interface by a bonding action, thus obtaining a composite membrane of the metal nano material and the biomolecules; and finally, extracting the biomolecule-metal composite monodisperse membrane on the liquid-liquid interface by a langmuir technology or directly by a solid substrate. The method for fixing biomolecules provided by the invention has a simple preparation process for forming the biomolecule-metal nano composite material, and has the characteristic that the biomolecules are dispersed in order in a single layer and have good stability.

Description

technical field [0001] The invention relates to a preparation method of a biosensor identification element, in particular to a method for immobilizing biomolecules. Background technique [0002] Biosensors use biological substances (such as enzymes, proteins, DNA, antibodies, antigens, biofilms, microorganisms, cells, etc.) Devices for detection and monitoring of substances. [0003] Biosensors are sensitive to biological substances. They use biologically active units (such as enzymes, antibodies, nucleic acids, cells, etc.) as biologically sensitive units, and are highly selective for target analytes. Its recognition elements are usually immobilized biomolecules (biosensitive materials, including enzymes, antibodies, antigens, microorganisms, cells, tissues, nucleic acids and other biologically active substances) immobilized on the substrate material. In order to develop low-cost, high-sensitivity, good-selectivity and long-life biosensors, immobilization technology has b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D71/82B01D69/12B01D67/00
Inventor 戴志晖刘倩包建春韩敏
Owner NANJING NORMAL UNIVERSITY
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