Method for separation and purification of streptavidin by means of nano magnetic particles
A nano-magnetic and particle separation technology, applied in chemical instruments and methods, desipeptides, peptides, etc., can solve problems such as high cost, complicated operation, and long time
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Embodiment 1
[0021] Weigh FeSO 4 ·7H 2 O (analytically pure) 4.12 grams, FeCl 3 ·6H 2 O (analytical pure) 8.03 grams, add double distilled water 150mL, dissolve under the protection of nitrogen, and add 300mL 5MNaOH (analytical pure) solution, stir vigorously, 80 ℃ of reaction 60 minutes, prepared magnetic nano Fe 3 o 4， Under an external magnetic field, wash with deionized water to neutrality, and then wash with methanol. In the methanol solution (adjust pH=3 with acetic acid), add 3.5 grams of silylating agent γ-aminopropyltriethoxysilane (Si (OC 2 h 5 ) 3 C 3 h 7 NH 2 ), reacted in a water bath at 60° C. for 4 hours, cooled, and washed with deionized water to obtain magnetic particles with amino groups on their surfaces. The particle size is detected by transmission electron microscopy, and the particle size is 20nm; the particles after surface silanization modification are detected by infrared, and the amino group (-NH 2 ) characteristic absorption at 3300cm -1 , N-H bendin...
Embodiment 2
[0025] Weigh FeSO4 ·7H 2 O (analytically pure) 4.12 grams, FeCl 3 ·6H 2 O (analytical pure) 8.03 grams, add double distilled water 150mL, dissolve under the protection of nitrogen, and add 300mL 5MNaOH (analytical pure) solution, stir vigorously, 80 ℃ of reaction 60 minutes, prepared magnetic nano Fe 3 o 4 , under an external magnetic field, wash with deionized water to neutrality, then wash with methanol, in the methanol solution (adjust pH=3 with acetic acid), add 3.5 grams of silylating agent γ-aminopropyltriethoxysilane ( Si(OC 2 h 5 ) 3 C 3 h 7 NH 2 ), reacted in a water bath at 60° C. for 4 hours, cooled, and washed with deionized water to obtain magnetic particles with amino groups on their surfaces. The size of the particles was detected by transmission electron microscopy, and the particle size was 20nm; the particles after surface silanization modification were detected by infrared to obtain amino (-NH 2 ) characteristic absorption at 3300cm -1 , N-H bendi...
Embodiment 3
[0029] Add 15.0 g of oleic acid-coated magnetic fluid, 32 mL of a mixture of styrene and 0.84 mL of divinylbenzene, and 240 mL of an aqueous solution of 5% sodium dodecyl sulfate into a 500 mL beaker. After stirring for 1 hour, add 0.6g of azodicyanovaleric acid, then dispersed at a high speed of 4000r / min for 30min, and transferred the mixed solution to a four-neck flask equipped with a stirrer, a condenser tube and a nitrogen inlet. Under the protection of nitrogen, the reaction was stirred at 80°C for 12 hours, and the emulsion was brown, which was separated and washed by a magnetic field, and collected into a grinding bottle. The size of the particle is detected by transmission electron microscopy, and the particle is at 300nm; the surface of the microsphere is detected by infrared with a carboxyl functional group, and the characteristic absorption of (-C=O) in the carboxyl group is at 1720cm -1 , (-C-O) characteristic absorption of stretching vibration at 1315cm -1 , (-O...
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