RU2011144738A - METHOD FOR PRODUCING AND CLEANING PLACENTAL ALKALINE PHOSPHATASE - Google Patents
METHOD FOR PRODUCING AND CLEANING PLACENTAL ALKALINE PHOSPHATASE Download PDFInfo
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- RU2011144738A RU2011144738A RU2011144738/15A RU2011144738A RU2011144738A RU 2011144738 A RU2011144738 A RU 2011144738A RU 2011144738/15 A RU2011144738/15 A RU 2011144738/15A RU 2011144738 A RU2011144738 A RU 2011144738A RU 2011144738 A RU2011144738 A RU 2011144738A
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- enzyme
- extract
- subjected
- heat treatment
- ammonium sulfate
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- Enzymes And Modification Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Способ получения и очистки плацентарной щелочной фосфатазы, заключающийся в экстрагировании фермента из сырья, термообработки экстракта, фракционировании белков сульфатом аммония и последующей очистки фермента методом препаративного электрофореза, отличающийся тем, что после экстрагирования фермента из плаценты 3М хлоридом калия и прогревания первичного экстракта в течение 10 мин при 65°C, проводят повторную термообработку экстракта в присутствии сульфата кадмия 0,14%-ной концентрации в течение 1-3 мин при 65°C с последующим охлаждением колбы под струей воды, добавляют в экстракт сульфат аммония до 30% насыщения, отделяют осадок центрифугированием, надосадочную жидкость, минуя стадию диализа, подвергают гидрофобной хроматографии на колонке с фенил-сефарозой, проводят элюцию фермента с колонки 0,02М трис-глициновым электродным буфером рН 8,6, а ферментсодержащий элюат подвергают препаративному электрофорезу в 7%-ном полиакриламидном геле.The method of obtaining and purification of placental alkaline phosphatase, which consists in extracting the enzyme from raw materials, heat treatment of the extract, fractionation of proteins with ammonium sulfate and subsequent purification of the enzyme by preparative electrophoresis, characterized in that after extraction of the enzyme from the placenta with 3M potassium chloride and heating of the primary extract for 10 min at 65 ° C, repeated heat treatment of the extract is carried out in the presence of cadmium sulfate of 0.14% concentration for 1-3 minutes at 65 ° C, followed by cooling to foreheads under a stream of water, ammonium sulfate is added to the extract to 30% saturation, the precipitate is separated by centrifugation, the supernatant, bypassing the dialysis stage, is subjected to hydrophobic chromatography on a phenyl-sepharose column, the enzyme is eluted from the column with 0.02 M Tris-glycine electrode pH buffer 8.6, and the enzyme-containing eluate is subjected to preparative electrophoresis in 7% polyacrylamide gel.
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Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
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RU2011144738/15A RU2492868C2 (en) | 2011-11-03 | 2011-11-03 | Method for preparing and cleaning placental alkaline phosphatase |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
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RU2011144738/15A RU2492868C2 (en) | 2011-11-03 | 2011-11-03 | Method for preparing and cleaning placental alkaline phosphatase |
Publications (2)
Publication Number | Publication Date |
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RU2011144738A true RU2011144738A (en) | 2013-05-10 |
RU2492868C2 RU2492868C2 (en) | 2013-09-20 |
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Family Applications (1)
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RU2011144738/15A RU2492868C2 (en) | 2011-11-03 | 2011-11-03 | Method for preparing and cleaning placental alkaline phosphatase |
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RU (1) | RU2492868C2 (en) |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0712434B2 (en) * | 1987-11-27 | 1995-02-15 | 日東化学工業株式会社 | Method for producing oxide catalyst containing phosphorus and antimony |
SU1576563A1 (en) * | 1988-03-15 | 1990-07-07 | Научно-производственное объединение "Фермент" | Method of obtaining alkaline phosphatase |
RU2016899C1 (en) * | 1991-06-19 | 1994-07-30 | Научно-исследовательский институт текстильных материалов | Method of alkaline phosphatase preparing |
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2011
- 2011-11-03 RU RU2011144738/15A patent/RU2492868C2/en not_active IP Right Cessation
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RU2492868C2 (en) | 2013-09-20 |
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MM4A | The patent is invalid due to non-payment of fees |
Effective date: 20131104 |