Amido modified magnetic nano-particle and method for preparing immune magnetic nano-separation agent

Abstract

The invention discloses a preparation method of magnetic nano-particles modified by aminogen. The preparation method comprises the steps that: (1) the alcoholysis of the magnetic nano-particles is carried out in a mixed solvent of toluene and low class monohydric alcohol, and surfaces of the magnetic nano-particles are coated by silica gel; the magnetic nano-particles coated by the silica gel areobtained; (2) the magnetic nano-particles coated by the silica gel react with a silane coupling agent, and the magnetic nano-particles coated by the silica gel with surfaces to modify an amino functional group are made. The invention also discloses a preparation method of an immunity magnetic nano separation agent which comprises the preparation method. The preparation method of the invention hasthe advantages of stable and moderate reaction system, simple and controllable operation and cheap agent material cost, and the waste reaction solution is easy to treat, and does not pollute environment. The preparation method is adapted to the industrial production. The magnetic nano-particles modified by aminogen has the advantages of small particle size, narrow distribution and good dispersibility, and the immunity magnetic nano separation agent has the advantages of high antibody combining rate, low non-specific binding rate, rapid magnetic separation, etc.

Description

technical field

[0001] The invention relates to an amino-modified magnetic nano particle suitable for industrialization and a preparation method of an immunomagnetic nano-separation reagent prepared therefrom. Background technique

[0002] In the late 1970s, magnetic separation technology began to be applied in the field of biology, and has achieved remarkable research results in cytology, molecular biology, biochemistry and biomedicine.

[0003] Magnetic separation technology uses magnetic particles as a carrier to coat biomolecules such as enzymes, proteins, peptides, antibodies, and antigens. Under the directional control of an external magnetic field, through operations such as affinity adsorption, cleaning, and desorption, complex biological molecules can be separated in one step. The target molecule is separated in the system, which has many advantages such as simple and convenient magnetic separation, high specificity and high sensitivity of affinity adsorption, so it...

Examples

Embodiment 1

[0035] 1) Prepare 1250ml0.5M NaOH and 125ml1M FeCl respectively 3 , 125ml0.5M FeCl 2 and 125ml0.4M HCl solution, NaOH solution was placed in a three-necked flask, under N 2 Heating the constant temperature water bath to 60°C under protective conditions, the prepared FeCl 3 , FeCl 2 After mixing with HCl solution, it was quickly added to a three-necked flask, stirred at 50 rpm for 15 min, and then cooled to room temperature. Washed 5 times with deionized water and methanol successively, the resulting product Fe 3 o 4 Magnetic nanoparticles were dispersed in methanol. The TEM image of the obtained product is shown in Figure 1(a); and the magnetic properties of the product are detected, as shown in the hysteresis curve in Figure 2.

[0036] 2) The product prepared in the previous step (Fe dispersed in methanol 3 o 4 Magnetic nanoparticles, Fe 3 o 4 Magnetic nanoparticles (20g) were placed in a flask, a total of 1000ml of toluene and methanol were added at a volume ratio...

Embodiment 2

[0041] 1) Prepare 1250ml0.5M KOH and 125ml1M Fe respectively 2 (SO 4 ) 3 , 125ml0.5M FeSO 4 and 125ml of 0.4M HCl solution, the KOH solution was placed in a three-necked flask, heated to 70°C at a constant temperature under the protection of argon (Ar), and the prepared Fe 2 (SO 4 ) 3 , FeSO 4 After mixing with HCl solution, it was quickly added into a three-necked flask, stirred vigorously at 100 rpm for 20 min, and then cooled to room temperature. Washed several times with deionized water and ethanol successively, the obtained product Fe 3 o 4 Magnetic nanoparticles are dispersed in ethanol.

[0042] 2) Put 20 g of magnetic nanoparticles prepared in the previous step in a flask, add 1050 ml of toluene and ethanol at a volume ratio of 1:15, add 10 ml of sodium silicate, and raise the temperature of the flask to 100 ° C, and pass in Ar for protection to Stir vigorously at 100 rpm, reflux for 24 hours, and wash with toluene several times to obtain Fe coated with silica...

Embodiment 3

[0047] 1) Prepare 1250ml0.5M KOH and 125ml1M Fe respectively 2 (SO 4 ) 3 , 125ml0.5M FeSO 4 and 125ml0.4M HCl solution, put the KOH solution in a three-necked flask, under N 2 Heating at constant temperature to 90°C under protective conditions, the prepared Fe 2 (SO 4 ) 3 , FeSO 4 After mixing with HCl solution, it was quickly added into a three-necked flask, stirred vigorously at 80 rpm for 30 min, and then cooled to room temperature. Wash several times with deionized water and methanol successively, and the obtained product Fe 3 o 4 Magnetic nanoparticles are dispersed in methanol.

[0048] 2) Put 20g of the magnetic nanoparticles prepared in the previous step in a flask, add 1000ml of toluene and methanol at a volume ratio of 1:30, add 200ml of tetraethyl orthosilicate, and raise the temperature of the flask to 120°C, and pass in N2 to carry out Protected, stirred vigorously at 80rpm, refluxed for 30h, washed several times with toluene to obtain Fe coated with sil...

technical field

[0001] The invention relates to an amino-modified magnetic nano particle suitable for industrialization and a preparation method of an immunomagnetic nano-separation reagent prepared therefrom. Background technique

[0002] In the late 1970s, magnetic separation technology began to be applied in the field of biology, and has achieved remarkable research results in cytology, molecular biology, biochemistry and biomedicine.

[0003] Magnetic separation technology uses magnetic particles as a carrier to coat biomolecules such as enzymes, proteins, peptides, antibodies, and antigens. Under the directional control of an external magnetic field, through operations such as affinity adsorption, cleaning, and desorption, complex biological molecules can be separated in one step. The target molecule is separated in the system, which has many advantages such as simple and convenient magnetic separation, high specificity and high sensitivity of affinity adsorption, so it...

Examples

Embodiment 1

[0035] 1) Prepare 1250ml0.5M NaOH and 125ml1M FeCl respectively 3 , 125ml0.5M FeCl 2 and 125ml0.4M HCl solution, NaOH solution was placed in a three-necked flask, under N 2 Heating the constant temperature water bath to 60°C under protective conditions, the prepared FeCl 3 , FeCl 2 After mixing with HCl solution, it was quickly added to a three-necked flask, stirred at 50 rpm for 15 min, and then cooled to room temperature. Washed 5 times with deionized water and methanol successively, the resulting product Fe 3 o 4 Magnetic nanoparticles were dispersed in methanol. The TEM image of the obtained product is shown in Figure 1(a); and the magnetic properties of the product are detected, as shown in the hysteresis curve in Figure 2.

[0036] 2) The product prepared in the previous step (Fe dispersed in methanol 3 o 4 Magnetic nanoparticles, Fe 3 o 4 Magnetic nanoparticles (20g) were placed in a flask, a total of 1000ml of toluene and methanol were added at a volume ratio...

Embodiment 2

[0041] 1) Prepare 1250ml0.5M KOH and 125ml1M Fe respectively 2 (SO 4 ) 3 , 125ml0.5M FeSO 4 and 125ml of 0.4M HCl solution, the KOH solution was placed in a three-necked flask, heated to 70°C at a constant temperature under the protection of argon (Ar), and the prepared Fe 2 (SO 4 ) 3 , FeSO 4 After mixing with HCl solution, it was quickly added into a three-necked flask, stirred vigorously at 100 rpm for 20 min, and then cooled to room temperature. Washed several times with deionized water and ethanol successively, the obtained product Fe 3 o 4 Magnetic nanoparticles are dispersed in ethanol.

[0042] 2) Put 20 g of magnetic nanoparticles prepared in the previous step in a flask, add 1050 ml of toluene and ethanol at a volume ratio of 1:15, add 10 ml of sodium silicate, and raise the temperature of the flask to 100 ° C, and pass in Ar for protection to Stir vigorously at 100 rpm, reflux for 24 hours, and wash with toluene several times to obtain Fe coated with silica...

Embodiment 3

[0047] 1) Prepare 1250ml0.5M KOH and 125ml1M Fe respectively 2 (SO 4 ) 3 , 125ml0.5M FeSO 4 and 125ml0.4M HCl solution, put the KOH solution in a three-necked flask, under N 2 Heating at constant temperature to 90°C under protective conditions, the prepared Fe 2 (SO 4 ) 3 , FeSO 4 After mixing with HCl solution, it was quickly added into a three-necked flask, stirred vigorously at 80 rpm for 30 min, and then cooled to room temperature. Wash several times with deionized water and methanol successively, and the obtained product Fe 3 o 4 Magnetic nanoparticles are dispersed in methanol.

[0048] 2) Put 20g of the magnetic nanoparticles prepared in the previous step in a flask, add 1000ml of toluene and methanol at a volume ratio of 1:30, add 200ml of tetraethyl orthosilicate, and raise the temperature of the flask to 120°C, and pass in N2 to carry out Protected, stirred vigorously at 80rpm, refluxed for 30h, washed several times with toluene to obtain Fe coated with sil...

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