RU96113901A - CHROMATOGRAPHIC METHOD FOR ISOLATING ALPHA-FETOPROTEIN - Google Patents

CHROMATOGRAPHIC METHOD FOR ISOLATING ALPHA-FETOPROTEIN

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Publication number
RU96113901A
RU96113901A RU96113901/25A RU96113901A RU96113901A RU 96113901 A RU96113901 A RU 96113901A RU 96113901/25 A RU96113901/25 A RU 96113901/25A RU 96113901 A RU96113901 A RU 96113901A RU 96113901 A RU96113901 A RU 96113901A
Authority
RU
Russia
Prior art keywords
solution
stage
fetoprotein
column
alpha
Prior art date
Application number
RU96113901/25A
Other languages
Russian (ru)
Other versions
RU2094077C1 (en
Inventor
Д.В. Кулаев
С.М. Насибов
С.С. Маркин
Ю.Г. Бобков
М.П. Семенов
Original Assignee
Акционерное общество закрытого типа "Фосфосорб"
Filing date
Publication date
Application filed by Акционерное общество закрытого типа "Фосфосорб" filed Critical Акционерное общество закрытого типа "Фосфосорб"
Priority to RU96113901A priority Critical patent/RU2094077C1/en
Priority claimed from RU96113901A external-priority patent/RU2094077C1/en
Application granted granted Critical
Publication of RU2094077C1 publication Critical patent/RU2094077C1/en
Publication of RU96113901A publication Critical patent/RU96113901A/en

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Claims (4)

1. Хроматографический способ выделения альфа-фетопротеина, включающий предварительную очистку исходного биологического фетального материала и многостадийную колоночную хроматографию, отличающийся тем, что предварительную очистку проводят путем осаждения раствором сульфата аммония, а хроматографию ведут в четыре стадии, на первой стадии - на колонке, заполненной цианбромированной сефарозой, иммобилизованной поликлональными антителами животных против альфа-фетопротеина человека с элюацией альфа-фетопротеина раствором карбоната натрия, на второй стадии - на колонке с ионообменной диэтиламиноэтил сефарозой уравновешенной натрий-фосфатным буфером при элюации раствором ацетатного буфера и нейтрализацией элюата раствором гидросида натрия, на третьей стадии - на колонке, заполненной цианбромированной сефарозой, иммобилизованной антителами животных против белков нормальной сыворотки человека при элюации раствором карбоната натрия, а на четвертой стадии - вновь на колонке с ионообменной диэтиламиноэтил сефарозой, уравновешенной натрий-фосфатным буфером при элюации раствором хлорида натрия, содержащего натрий-фосфатный буфер и сахарозу.1. A chromatographic method for the isolation of alpha-fetoprotein, including preliminary purification of the initial biological fetal material and multi-stage column chromatography, characterized in that the preliminary purification is carried out by precipitation with a solution of ammonium sulfate, and the chromatography is carried out in four stages, in the first stage - on a column filled with cyanogen bromide sepharose immobilized by animal polyclonal antibodies against human alpha-fetoprotein with elution of alpha-fetoprotein with a carbonate solution on ria, in the second stage, on a column with ion-exchange diethylaminoethyl sepharose balanced with sodium phosphate buffer, eluting with an acetate buffer solution and neutralizing the eluate with sodium hydroxide solution, in the third stage, on a column filled with cyanogen brominated sepharose immobilized with animal antibodies against normal protein a solution of sodium carbonate, and in the fourth stage, again on a column with ion-exchange diethylaminoethyl sepharose, balanced with sodium phosphate buffer during elution a solution of sodium chloride containing sodium phosphate buffer and sucrose. 2. Способ по п. 1, отличающийся тем, что после колоночной хроматографии продукт замораживают и сушат лиофильно. 2. The method according to p. 1, characterized in that after column chromatography the product is frozen and freeze-dried. 3. Способ по п. 1, отличающийся тем, что в качестве биологического фетального материала используют абортальную или платентарную сыворотку или пуповинную кровь. 3. The method according to p. 1, characterized in that as a biological fetal material using abortion or platent serum or cord blood. 4. Способ по п. 1, отличающийся тем, что в качестве антител животных используют антитела кроликов, овец, коз, мышей. 4. The method according to p. 1, characterized in that the antibodies of animals use antibodies of rabbits, sheep, goats, mice.
RU96113901A 1996-07-23 1996-07-23 Chromatographic method for isolation of alfa-fetoprotein RU2094077C1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
RU96113901A RU2094077C1 (en) 1996-07-23 1996-07-23 Chromatographic method for isolation of alfa-fetoprotein

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
RU96113901A RU2094077C1 (en) 1996-07-23 1996-07-23 Chromatographic method for isolation of alfa-fetoprotein

Publications (2)

Publication Number Publication Date
RU2094077C1 RU2094077C1 (en) 1997-10-27
RU96113901A true RU96113901A (en) 1998-01-20

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Family Applications (1)

Application Number Title Priority Date Filing Date
RU96113901A RU2094077C1 (en) 1996-07-23 1996-07-23 Chromatographic method for isolation of alfa-fetoprotein

Country Status (1)

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RU (1) RU2094077C1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SI1869065T1 (en) * 2005-03-11 2020-08-31 Wyeth Llc A method of weak partitioning chromatography

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