Purification method, extract and preparation of cobra venom neurotoxin
A technology for cobra venom and neurotoxin, which is applied to the preparation methods of peptides, nervous system diseases, chemical instruments and methods, etc., can solve the problems of difficult quality control, low purity, long production cycle, etc., and achieves improved separation and simplified extraction process. , the effect of saving time and cost
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Embodiment 1
[0094] Embodiment 1: the preparation of cobra venom neurotoxin
[0095] 1) Dissolve 100g of cobra venom powder in pH7.9, 0.01M phosphate buffer to 280ml, dissolve under stirring, centrifuge at 3000r / min for 10min, take the supernatant, pass the supernatant through 0.22μm membrane;
[0096] 2) The obtained supernatant is subjected to ultrafiltration and exchanged for a separation buffer, using an ultrafiltration membrane with a molecular weight cut-off of 10KDa;
[0097] 3) The obtained snake venom is loaded onto the CM-cellulose chromatography column with pH7.9, 0.01M phosphate buffer balance, and is eluted with phosphate buffer, and the elution flow rate is 40ml / h. The eluate was collected by an automatic fraction collector and detected at 280 nm by a UV spectrophotometer.
[0098] 4) When the absorbance of the eluate drops below 0.2, elute with a phosphate buffer containing 0.04mol / L Nacl, and when the absorbance of the eluate drops below 0.2, elute with a phosphate buffer...
Embodiment 2
[0100] Embodiment 2: the preparation of cobra venom neurotoxin extract
[0101] 1) Dissolve 100g of cobra venom powder in pH 8.0, 0.01M phosphate buffer to 280ml, dissolve under stirring, centrifuge at 3000r / min for 10min, take the supernatant, pass the supernatant through 0.22μm membrane;
[0102] 2) The obtained supernatant is subjected to ultrafiltration and exchanged for a separation buffer, using an ultrafiltration membrane with a molecular weight cut-off of 10KDa;
[0103] 3) The obtained snake venom is loaded onto the CM-cellulose chromatography column with pH8.0, 0.01M phosphate buffer balance, and is eluted with phosphate buffer, and the elution flow rate is 50ml / h. The eluate was collected by an automatic fraction collector and detected at 280 nm by a UV spectrophotometer.
[0104] 4) When the absorbance of the eluent drops below 0.2, elute with a phosphate buffer containing 0.05 mol / L Nacl. After the absorbance of the eluate drops below 0.2, take the part with th...
Embodiment 3
[0106] Embodiment 3: the preparation of cobra venom neurotoxin extract
[0107] 1) Dissolve 100g of cobra venom powder in pH7.8, 0.01M phosphate buffer to 280ml, dissolve under stirring, centrifuge at 3000r / min for 10min, take the supernatant, pass the supernatant through 0.22μm membrane;
[0108] 2) The obtained supernatant is subjected to ultrafiltration and exchanged for a separation buffer, using an ultrafiltration membrane with a molecular weight cut-off of 10KDa;
[0109] 3) The obtained snake venom is loaded onto the CM-cellulose chromatographic column that has been equilibrated with pH7.8, 0.01M phosphate buffer, and is eluted with phosphate buffer, and the elution flow rate is 30ml / h. The eluate was collected by an automatic fraction collector and detected at 280 nm by a UV spectrophotometer.
[0110] 4) When the absorbance of the eluent drops below 0.2, elute with a phosphate buffer containing 0.03mol / L Nacl, and when the absorbance of the eluate drops below 0.2, e...
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