SU1187835A1 - Method of affine chromatography of biologically active substances - Google Patents
Method of affine chromatography of biologically active substances Download PDFInfo
- Publication number
- SU1187835A1 SU1187835A1 SU833650787A SU3650787A SU1187835A1 SU 1187835 A1 SU1187835 A1 SU 1187835A1 SU 833650787 A SU833650787 A SU 833650787A SU 3650787 A SU3650787 A SU 3650787A SU 1187835 A1 SU1187835 A1 SU 1187835A1
- Authority
- SU
- USSR - Soviet Union
- Prior art keywords
- molecular weight
- biologically active
- active substances
- sorbent
- chromatographed
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 9
- 239000013543 active substance Substances 0.000 title claims 2
- 238000004587 chromatography analysis Methods 0.000 title abstract description 3
- PXFBZOLANLWPMH-UHFFFAOYSA-N 16-Epiaffinine Natural products C1C(C2=CC=CC=C2N2)=C2C(=O)CC2C(=CC)CN(C)C1C2CO PXFBZOLANLWPMH-UHFFFAOYSA-N 0.000 title abstract 2
- 150000003839 salts Chemical class 0.000 claims abstract description 4
- 239000002594 sorbent Substances 0.000 claims abstract 4
- 230000007717 exclusion Effects 0.000 claims abstract 2
- 239000003446 ligand Substances 0.000 claims abstract 2
- 101000608750 Arachis hypogaea Alpha-methyl-mannoside-specific lectin Proteins 0.000 claims 1
- 101000771730 Tropidolaemus wagleri Waglerin-3 Proteins 0.000 claims 1
- 238000001042 affinity chromatography Methods 0.000 claims 1
- 102000009027 Albumins Human genes 0.000 description 4
- 108010088751 Albumins Proteins 0.000 description 4
- 239000000872 buffer Substances 0.000 description 3
- 239000007974 sodium acetate buffer Substances 0.000 description 3
- 102000016911 Deoxyribonucleases Human genes 0.000 description 2
- 108010053770 Deoxyribonucleases Proteins 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
1. СПОСОБ АФФИННОЙ ХРОМАТОГРАФИИ виологачЕски АКТИВНЫХ ввищств отличающийс тем, что, с целью ускорени способа, комплекс лиганд-аффинант хроматографируют на сорбенте при, градиенте концентрации соли 0,02-1,5 М. 2. Способ по П.1, отличающийс тем, что, с целью повьшени степени чистоты, целевой продукт хроматографируют на сорбенте, у которого молекул рна масса исключени равна молекул рной массе лиганда, а нижний предел разрешени равен молекул рной массе аффинанта.1. METHOD OF AFFINE CHROMATOGRAPHY viologuSkI ACTIVE vizvchstv characterized in that, in order to accelerate the method, the complex ligand-affinant is chromatographed on a sorbent at a salt concentration gradient of 0.02-1.5 M. that, in order to increase the purity, the target product is chromatographed on a sorbent, in which the exclusion molecular weight is equal to the molecular weight of the ligand, and the lower resolution limit is equal to the molecular weight of the affinant.
Description
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Изобретение относитс к биохимии, в частности к способам получени и ОЧР1СТКИ макромолекул из микробиологического и другого вида сырь .The invention relates to biochemistry, in particular, to methods for the production and EFFICIENCY of macromolecules from microbiological and other types of raw materials.
Целью изобретени вл етс ускорение способа, повьппение степени чистоты целевого продукта.The aim of the invention is to accelerate the process by increasing the purity of the target product.
П р и м е р. 1. Колонку 2,5 Я 95см с Сфероиом 1000 (пределы разрешени по мол.м. дл белков 800000-5000000) уравновешивали 1,5 л 20 мМ Трис-НС1 буфера, рН 7,5, содержащего 1,5 М КС1 Затем в колонку вводили линейный градиент концентрации КС1 от 1,5 до 0,1 М (160 мл) со скоростью 70 мл/ч. 3 мл раствора альбумина (20 мг) в . 0 мМ Трис-НС1 буфере рН 7,5, содержащем О,1 М КС1 и 60 мг голубого декстрана (мол.м. 2000000), наносили на колонку вслед за градиентом соли и промывали 10 мМ Трис-НС1 буфером, рН 7,5 - 0,1 М КС1. Профиль элюции альбумина показан на фиг.1. Дл сравнени показан профиль элюции альбумина , полученный при хроматографии его без голубого декстрана. На фиг.2 показана денситограмма, полученна после электрофореза альбумина до и после очистки указанным способом.PRI me R. 1. A column of 2.5 I 95 cm with Spheroi 1000 (resolution mol. M. For proteins 800000-5000000) was equilibrated with 1.5 l of 20 mM Tris-HCl buffer, pH 7.5, containing 1.5 M KCl. Then in the column was injected with a linear concentration gradient KC1 from 1.5 to 0.1 M (160 ml) at a rate of 70 ml / h. 3 ml of albumin (20 mg) c. 0 mM Tris-HC1 buffer pH 7.5, containing O, 1 M KCl and 60 mg of blue dextran (mol.m. 2,000,000) were applied onto the column following the salt gradient and washed with 10 mM Tris-HC1 buffer, pH 7.5 - 0.1 M KS1. The albumin elution profile is shown in FIG. For comparison, the elution profile of albumin obtained by chromatography without blue dextran is shown. Figure 2 shows a densitogram obtained after electrophoresis of albumin before and after purification by this method.
Пример 2. Колонку 1,6jf 40 см с Toyopearl - 75Г (пределы разрешени по мол.м. дл белков 500000 50000000 ) уравновешивали 0,3 л 0,05 М натрий-ацетатным буфером (рН 4,0), содержащим 1,5 М КС1, со скоростью 60 мл/ч. Затем в колонку вводили градиент концентрации КС1 от 1,5 до 0,05 М (30 мл). 0,8 мл смеси, содержащей 3 мг ДНК тимуса теленка и 3,0 мг ДНКазы 1 в 0,05 М натрий-ацетатном буфере (рН 4,0), наносили на колонку вслед за градиентом соли и промывали 0,05 М натрий-ацетатным буфером (рН 4,0), содержащим 0,05 М КС1. Первой с колонки элюировалась ДНК, затем ДНКаза 1 (2,48 мг) и следом примеси, загр зн ющие препарат фермента.Example 2. A column of 1.6 jf 40 cm with Toyopearl-75G (resolution limits in mol.m. for proteins 500000 50000000) was equilibrated with 0.3 l of 0.05 M sodium acetate buffer (pH 4.0) containing 1.5 M KS1, with a speed of 60 ml / h. Then, a concentration gradient of KCl was introduced into the column from 1.5 to 0.05 M (30 ml). 0.8 ml of a mixture containing 3 mg of calf thymus DNA and 3.0 mg DNase 1 in 0.05 M sodium acetate buffer (pH 4.0) was applied onto the column after a salt gradient and washed with 0.05 M sodium acetate buffer (pH 4.0) containing 0.05 M KCl. DNA was first eluted from the column, then DNase 1 (2.48 mg) and a trace of impurity contaminating the enzyme preparation.
« vНапрабление электродуореза“V
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU833650787A SU1187835A1 (en) | 1983-10-10 | 1983-10-10 | Method of affine chromatography of biologically active substances |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU833650787A SU1187835A1 (en) | 1983-10-10 | 1983-10-10 | Method of affine chromatography of biologically active substances |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SU1187835A1 true SU1187835A1 (en) | 1985-10-30 |
Family
ID=21084876
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SU833650787A SU1187835A1 (en) | 1983-10-10 | 1983-10-10 | Method of affine chromatography of biologically active substances |
Country Status (1)
| Country | Link |
|---|---|
| SU (1) | SU1187835A1 (en) |
-
1983
- 1983-10-10 SU SU833650787A patent/SU1187835A1/en active
Non-Patent Citations (1)
| Title |
|---|
| J.Biochem., 1982, 91,р. 1411-1417. * |
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