Immunomagnetic nano particle cell separator, and its preparing method and use
A magnetic nanoparticle and cell separator technology, applied in biochemical instruments, biochemical equipment and methods, bioreactor/fermenter combination, etc., can solve the problem of inconvenient separation of CD34+ cells, and there is no report on the structure of magnetic nanoparticle cell separator And other issues
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[0030] The method for preparing an immunomagnetic nanoparticle cell separator of the present invention includes the following steps:
[0031] (1) Preparation of magnetic nanoparticles
[0032] Separately prepare FeSO with double distilled water 4 .7H 2 O and FeCl 3 .6H 2 O mixed solution and NaOH solution. Fe in the mixed solution of iron salt 2+ The ion concentration is 0.1-0.2mol / l, Fe 3+ The concentration of ions is 0.1-0.3 mol / l, and the concentration of NaOH solution is 2-3 mol / l. Under vigorous stirring, a NaOH solution whose volume is half of the volume of the mixed salt solution is slowly added dropwise to the mixed salt solution. The obtained solid precipitate was aged for 12h at 40℃-60℃, washed with double distilled water several times, filtered and then dried at 40℃-80℃ for 24h, then ground in an agate mortar Get the product.
[0033] (2) Silica in γ-Fe 2 O 3 Surface modification
[0034] TritonX-100, n-hexanol and cyclohexane are uniformly mixed in a ratio of 1:(1-3):(...
Embodiment 1
[0049] Method for preparing core layer of magnetic nano particles
[0050] Using improved chemical co-precipitation to prepare the core layer of magnetic particles, the specific method is as follows: separately prepare FeSO with double distilled water 4 .7H 2 O and FeCl 3 .6H 2 O mixed solution and NaOH solution. Fe in the mixed solution of iron salt 2+ The ion concentration is 0.1-0.2mol / l, Fe 3+ The concentration of ions is 0.1-0.3 mol / l, and the concentration of NaOH solution is 2-3 mol / l. Under vigorous stirring, a NaOH solution whose volume is half of the volume of the mixed salt solution is slowly added dropwise to the mixed salt solution. The obtained solid precipitate was aged for 12 hours at 40°C-60°C, washed with double distilled water several times, filtered and then dried at 40°C-80°C for 24 hours, and then ground in an agate mortar. Get the product.
Embodiment 2
[0052] Preparation method of core-shell type nucleic acid magnetic nano particles
[0053] TritonX-100, n-hexanol and cyclohexane are uniformly mixed in a ratio of 1:2:5 to form a transparent and stable microemulsion system. Put the above microemulsion system in ultrasonic for 30-60 minutes, and then add 0.5g of γ-Fe to it 2 O 3 , After ultrasonic treatment for 6 minutes, take out the upper layer and pour it into a three-necked flask, stir for 30 minutes to make it uniform. Dilute 1ml of a certain concentration of concentrated ammonia water with 2ml of double distilled water, slowly add it to the continuously stirred microemulsion after 30 minutes, and continue to stir for 30 minutes to make the ammonia water evenly dispersed in the microemulsion. After 1 hour, 1-3 ml of ethyl orthosilicate was added dropwise to the microemulsion while stirring continuously for 10 hours, and the temperature of the system was kept between 15-30°C. Add acetone to the system to precipitate the partic...
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