Placenta polypeptide preparation method
A placental polypeptide and placenta technology, which are applied to the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problems of low yield of polypeptides, and achieve the effects of simple preparation method, less equipment investment and low production cost.
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Embodiment 1
[0021] The placenta without hepatitis B, hepatitis C, AIDS, and syphilis virus is shredded and added to normal saline at a mass ratio of 1:2 for tissue homogenization; the resulting tissue homogenate is mixed with a compound enzyme at a mass ratio of 50:1, and the The complex enzyme is composed of trypsin and papain with a mass ratio of 1:1. It is hydrolyzed for 4 hours at 30°C and pH 5, and the centrifuge is collected by centrifugation at 5000r / min. The collected centrifugate is used as the pre-extraction aqueous phase. Hexanol and petroleum ether with a volume ratio of 1:3 are the organic phase, wherein the volume ratio of the pre-extracted organic phase and the aqueous phase is 2:1, and AOT and Span-60 with a mass ratio of 2:1 are used as a mixed surfactant. Extract at 40°C for 15 minutes, use 2mol / L KCl solution as the stripping aqueous phase, wherein the volume ratio of the post-extracted organic phase to the KCl solution aqueous phase is 2:1, strip at 35°C for 30 minutes,...
Embodiment 2
[0023] The placenta without hepatitis B, hepatitis C, AIDS, and syphilis virus is shredded and added to normal saline at a mass ratio of 1:2.5 for tissue homogenization; the resulting tissue homogenate is mixed with a compound enzyme at a mass ratio of 55:1, and the The compound enzyme is composed of five kinds of trypsin, pepsin, papain, bromelain and carboxypeptidase of the same quality. It is hydrolyzed at 35°C and pH 6 for 4.5h, centrifuged at 5000r / min to collect the centrifugate, and the collected The centrifuge liquid is used as the pre-extraction aqueous phase, and the hexanol and sherwood oil of 1:4 volume ratio are organic phases, wherein the organic phase and the aqueous phase volume ratio of pre-extraction are 2.5:1, with the AOT of 2.5:1 mass ratio and division Pan-60 is a mixed surfactant, extracted at 42°C for 16 minutes, using 2mol / L KCl solution as the stripping aqueous phase, wherein the volume ratio of the post-extracted organic phase to the KCl solution aque...
Embodiment 3
[0025] After the placenta without hepatitis B, hepatitis C, AIDS, and syphilis virus is shredded, add physiological saline in a mass ratio of 1:3 to carry out tissue homogenization; the resulting tissue homogenate is mixed with papain in a mass ratio of 60:1, and the ℃, pH 9 conditions of enzymatic hydrolysis for 5h, centrifugation at 5000r / min to collect the centrifugate, the collected centrifugate as the pre-extraction aqueous phase, with 1:5 volume ratio of hexanol and petroleum ether as the organic phase, of which the pre-extracted The volume ratio of the organic phase to the aqueous phase is 3:1, AOT and Span-60 with a mass ratio of 3:1 are used as mixed surfactants, extracted at 45°C for 20 minutes, and 2mol / L KCl solution is used as the back-extracted aqueous phase, The volume ratio of the post-extracted organic phase to the KCl solution aqueous phase is 3:1, back-extracted at 40°C for 40 minutes, and the crude extract of placental polypeptide and protease is obtained af...
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