DE2443119A1 - SELECTIVE REMOVAL OF ALBUMIN FROM BLOOD FLUIDS, AS WELL AS SUBSTANCES FOR CARRYING OUT THE PROCEDURE - Google Patents

Description

Selective removal of albumin from blood fluids as well as substances for performing the procedure

The invention relates to the selective fractionation of proteins in blood fluids.

It is known that blood fluids contain such as Plasma and serum, many proteins that one would like to isolate in a pure state. Isolation procedure for such proteins are known, but they all suffer have a common disadvantage, namely the presence of albumin, the main protein component in the Plasma. Apart from the fact that it is an undesirable contaminant in the processes mentioned, liked one often isolates pure albumin without simultaneous contamination by other proteins.

It is also known that certain proteins are bound very tightly to certain reactive dyes will. For example, dextran blue (Blue

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Dextran) firmly bound to lysozyme, ovalbumin and hemoglobin, if these are positively charged, as well regardless of the load of bovine serum albumin. Dextran blue is a dye commonly known as Procion Blue HBS and in which the chlorine atom of the triazine residue in the dye by polymeric dextran, which is attached to the Triazine residue is bound, is replaced (White and Jencks, Binding of Proteins to Blue Dextran, No. 43, Biol., Abstracts, 16th National Meeting, American Chemical Society, September 14, -18, 1970). However is Dextran blue soluble in water. Therefore, dextran blue would have to be coupled to a solid substrate from which the Albumin can be removed.

A method for protein purification by affinity chromatography is also known (J.Biol. Chem. 245 , 3059, (1970)). In this process, insoluble materials are produced by coupling certain large molecules containing amino groups to agarose, activating the agarose with cyanogen bromide and then reacting the activated agarose with the amino group-containing residue.

It is also known from J. Chromatogr., 6g, 209, (1972) that the condensation product of Cibacron Blue F3G-A, also known as Procion Blue HBS, with Dextran 2000, also known as Blue Dextran 2000 from Pharmacia, Uppsala, Sweden is known to have a high affinity for the enzyme phosphofructokinase. On the other hand, the condensation product of dextran showed 2000 with Cibacron Brilliant Blue FBR-P, a dye with an extremely similar structure, no affinity whatsoever versus phosphofructokinase. There was also no activity when using blue dextran Cyanogen bromide activated Sepharose coupled.

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The specificity of dyes for certain polypeptides is therefore unpredictable.

The object of the invention is therefore to develop a method by which albumin is selectively extracted from blood fluids can be removed without simultaneously removing other proteins that are in very small amounts may be present.

Another object of the invention is to create a method according to which the albumin can be removed so that the absorption capacity of the substrate does not suffer any loss and the albumin can be recovered so that it does not undergo denaturation.

It has been found that certain reactive dyes by coupling provide a substance for particular solid carrier phase, selectively to the aqueous albumin from virtually cell-free _fluids; such as plasma or serum, can be adsorbed. The adsorbed albumin can be separated again in high yield without denaturation from these substances, which in turn can be reused without loss of their adsorptive capacity. The carrier phases that can be used for this purpose are agarose, in particular Sepharose, polyacrylamides, in particular crosslinked polyacrylamide gel, and acrylic resins, in particular anion exchange resins, such as Amberlite IRC 45, which contain an amino group.

As already mentioned, these carrier phases are bound to certain reactive dyes CRD, whereby the dyes are all contained in the color index.

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The invention relates to the substances characterized in claim 1 and that in claim -4 marked procedures.

The adsorption of the albumin to the substance from the carrier phase and reactive dye from the to from it liberating liquid is expedient in a buffer with a molarity of at least 0.05 and carried out at a pH of 6.5 to 8.5. The suspension is expediently made from the substance Carrier phase and reactive dye placed on a chromatography column and the practically cell-free albumin-containing liquid was allowed to pass through the column. The eluate is practically free of albumin and can be further processed in any desired way.

The albumin is then expedient from the combination of carrier phase and reactive dye removed by eluting, preferably eluting a column. The elution media can be an aqueous urea solution, an aqueous / ethanolic solution of alkali phosphate, an aqueous solution of alkaline buffered Alkanecarboxylic acids with 2 to 10 carbon atoms or aqueous solutions of compounds from cations of Metals of group 2A of the periodic table and of potassium with pharmaceutically acceptable anions. Use of any of these eluants provides a carrier phase / reactive dye combination with renewed activity designed to remove additional amounts of albumin from additional albumin containing Liquids can be reused. However, if desired, pure, undenatured albumin To isolate, the two last-mentioned groups of solvents must be used. The albumin can then be isolated from them, for example by lyophilization, in the usual catfish.

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~ ⁵ ~ 2U3119

In one embodiment of the invention, agarose, preferably in the form of Sepharose, which can be in the form of packed Sepharose (for example of grade 2B, 4B or 6B) or crosslinked Sepharose (for example of grade 2B, 4B or 6B), can be used as the carrier phase . As mentioned above, the reactive dyes which can be used in accordance with the invention share certain structural features in common. Some of these dyes can be identified in more detail with the help of their general color index name, their color index number or their trade name. These dyes include the followingϊ CI Reactive Orange 2 (CI Constitution No. 17865 _ff also as Cibacron Orange GE or Procion Brilliant Orange HGRS bekanni ^, CI Reactive Red 4 (CICN 18105, also as Cibacron Brilliant Red 3B-A or Procion Brilliant Red H7BS), CI Reactive Brown 1 (CICN 26440, also known as Cibacron Brown 3GR-A or Procion Orange Brown HGS) j'CI Reactive Red 9 (CICN 17910, also known as Cibacron Scarlet 2G or Procion Scarlet H3GS.) From CI Reactive Red 16, also known as Cibacron Scarlet 4G-B, has a similar structure. This group also includes the anthraquinone colors CI Reactive Blue 2 (CICN 61211, also known as Cibacron Blue F3G-A or Procion Blue HBS and CI Reactive Blue 5 ( CICN 61210, also known as Cibacron Brilliant Blue FBR-P «,)

Processes for the production of all of the named dyes can be found in CoLCzech.Chem.Commune., 25 »2783 (1960).

Such compounds are also within the scope of the invention in which the terminal benzenesulfonic acid group is replaced by another substituent, such as an alkyl, phenyl *, alkylphehyl,

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Alkaxyphenyl group, hydrogen or the like. A typical representative of this group is CI Reactive Red (CICN 18159, cf. Coloristica Buletin Informativ, 4, (No. 11, 1968)
179-200.

All of the dyes mentioned can be coupled to the carrier phase substrates by heating in water at moderately elevated temperatures, expediently above a pH of 6.0. In a preferred embodiment of the process according to the invention, the mixtures are ^{heated to 75 to 95 °} C. with stirring, then cooled and washed to remove free dye.

The weight ratio of dye to carrier phase is not critical.

If too little dye is used, the effectiveness of the extraction process will be in view of the
reduced to a small extent existing adsorption sites. If too much dye is used, it will
The process is not only uneconomical, but steric disturbances also occur, which also lead to a reduction in the number of available adsorption sites for the albumin.

0.1 to 5 g of dye can be used per 4 g of carrier phase. Preferably, however, the use of _t 0 is 2 to 1 g of dye per 4 g of carrier phase. From a practical point of view, however, the carrier phase is usually not determined by weight, but rather by the volume of deposited carrier phase in aqueous suspension. In the case of Sepharose as the carrier phase, 4 g of Sepharose correspond to practically 100 cm of separated Sepharose.

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Of course, the weight to volume ratio will vary with the chemical nature of the carrier phase as well as the grain size determined by grades 2B, 4B and 6B of Sepharose for the generally available carrier phases is represented, but the ratios given are sufficiently accurate to be satisfactory To maintain workspace

In a preferred embodiment of the process according to the invention, the dye is dissolved or suspended in an amount of 0.1 to 2 g, preferably 0.2 to 1 g, in a small amount of water for the coupling (between 10 and 50 ml are generally sufficient ), and added to 100 ml of carrier phase in water (measured as the set carrier phase). The amount of water that is used to suspend the carrier phase is not of decisive importance? between 100 and 1000 ml have proven to be suitable. The mixture is stirred gently and warmed to below the boiling point of the water. A temperature range from 75 to 95 ^° C. has proven to be satisfactory. The pH of the solution is kept slightly above 6, expediently between 6 and 8, by adding either aqueous alkali or a buffer. The reaction time depends on the degree of dilution of the dye, the temperature and the agitation of the carrier phase. It is generally between about 5 and about 60 minutes. The coupling product (SP / CRD) is then separated from the aqueous phase - expediently by decanting - and washed to remove unreacted dye. It has proven to be useful to wash thoroughly with water at room temperature and then with a suitable salt, such as guanidine hydrochloride (5 molar), which removes any residues of unreacted dye. The use of guanidine hydrochloride solution

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however, it is not critical and other wash liquids can be used.

The coupling product purified in this way is then suspended in a buffer, expediently one with a pH of 6.5 to 8.5. The absorptive capacity of the product thus obtained for albumin naturally depends on the ratio of Dye to carrier phase. However, own products that are manufactured according to the regulations described have an absorptive capacity in the order of 30 mg albumin per ml deposited coupling product

Of course, different dyes react with different carrier phases at different speeds. In addition, care must be taken that the carrier phase is not physically changed, ie that it does not partially or completely melt during the coupling process. For example, Cibacron Blue F-3GA reacts very rapidly with Sepharose at 80 ⁰ C, while other dyes at this temperature couple very slowly with ordinary Sepharose. While the coupling ^{takes place rapidly at 90 °} C., the product produced, although usable, does not have the optimal physical properties. If, therefore, higher reaction temperatures are required, it is advantageous to use crosslinked carrier phases, such as, for example, crosslinked Sepharose. ·

Examples of further carrier phases that can be used in the method according to the invention, are polyacrylamide gels and acrylic resins containing amino groups, such as Amberlite® IRC 45.

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The dextran derivatives can also be used from Cibacron Blue F3GA and Cibacron Brilliant Blue FBR-P. In these dyes, the chlorine group is des Triazine residue replaced by an oxygen bond to a dextran molecule · The first mentioned dye supplies with Dextran 2000 the product Blue Dextran 2000®.

The processes mentioned for the preparation of the coupling products are based on the implementation of a. Halogen group on the triazine residue with a labile group on the carrier phase. Such a reaction cannot be carried out if this group has been replaced by a -0-dextran bond. Regardless of this, in those cases in which the dye has a labile amino group, such as the primary amino group on the anthraquinone residue of Cibacron Blue F3GA and Cibacron Brilliant Blue FBR-P, the coupling can be carried out using the above-mentioned cyanogen bromide method. In this process, the carrier phase is activated by treatment with aqueous alkaline cyanogen bromide and the appropriately buffered carrier phase is reacted with the dye. In this process, the product of dextran and dye is then added to a similar buffer of the activated carrier phase and the coupling is allowed to proceed. The coupling reaction can take place at temperatures between 0 ^° C. and room temperature, but it is preferably carried out ^{at about 4 ° C. for about 24 hours.} After this time has elapsed, the coupling product is successively mixed with aqueous sodium bicarbonate with a pH of about 9-5, concentrated aqueous urea (about 6 molar), water and a buffer with a high ion concentration and a pH of 6.5 to 8, 5 washed. A Tris-HCl / aqueous sodium chloride buffer is preferably used. The product made from sepharose, dextran and dye is made

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then suspended in fresh buffer and can be ^{stored at about 4 °} C. until it is used.

When performing albumin removal in practice it has proven to be useful to place the coupling product in a buffer with a high ion concentration to be introduced in a column. The above-mentioned Tris-HCl / sodium chloride buffer has proven to be particularly useful for this purpose proven. The high ion concentration of the buffer system is essential for the implementation of the The process is crucial because it prevents the coupling product from acting as a cation exchanger. It has It has been found to be expedient to add about 2 ml of albumin-containing plasma to about 10 ml of deposited coupling product use. The plasma is then passed through the column and the column with the said buffer washed. The elution was followed by uv adsorption of the eluate.

As mentioned above, the decisive factor in this method is the ion concentration of the buffer system. To be effective, the buffer must have at least a 0.05 molar ion concentration at pH from 6.5 to 8.5 and preferably about 8.0, with no upper limit to be observed.

The eluate obtained in this way is practically free of albumin and the remaining proteins can can be isolated from it in a known manner. Examination and comparison of the initial plasma and the eluate showed that apart from the albumin only removes small amounts of the other proteins, mainly lipoproteins will.

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The column can be broken by breaking the bond between see the albumin and the coupling product regenerated will. In those cases in which the preservation of the albumin is not necessary, an aqueous one has Urea solution proved to be useful for this. the Urea solution must be at least one molar, three molar Solutions have been used and any concentration up to saturation (around 8 mol / l) is satisfactory. A column volume of urea solution is sufficient to break the bonds, however usually 1 to 3 column volumes used.

In those cases in which it is desired to protect the albumin, an aqueous ethanolic solution of an alkali metal phosphate, expediently sodium phosphate, is used. The preferred volume ratio is 1 % 1, and the phosphate solution is 0.05 to 0.1 molar at a pH value of 2.0 to 3 and expediently 0.075 molar at a pH value of 2.4, measured before mixing.

It has also been found that albumin can be eluted in high purity using an aqueous solution of an alkaline earth cation and a pharmaceutically acceptable anion. Particularly suitable cations are Ca "*" ¹ "and Mg ⁺⁺ . K ⁺ , but not Na ⁺ , can also be used. Any strong pharmaceutically acceptable anion, suitably the chloride ion, can serve as the anion. The ion concentration of these eluents should preferably be at least 0 .05 "molar. Lower concentration tones cause tailing · Higher concentrations up to about 0.1 mol / l lead to faster elution and are acceptable, but the upper limits mentioned should not be viewed as limiting.

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It has proven to be expedient to buffer the eluant, expediently to a pH value between 6.0 and 9 and preferably from about 8. This is achieved by adding the eluent a buffer such as sodium bicarbonate or trisodium chloride is added. An ion concentration of about 0.05 "to 0.1 mol / l Tris hydrogen chloride in the eluent has been found to be suitable.

It has also been found that base-buffered alkanecarboxylic acids are suitable as eluants. Acids having 2 to 10 carbon atoms can be used, but those having 8 to 10 carbon atoms are preferred. Octanecarboxylic acid is particularly suitable as it is the stabilizer of choice for albumin. When using alkanecarboxylic acids, in particular octanecarboxylic acid, as eluent, is obtain a stabilized solution of pure albumin which, if desired, can be concentrated under reduced pressure or lyophilized, after which a pure buffered albumin is obtained.

Any buffer can be used that increases the pH of the solution to at least 6.5, the The range of 7.5 to 9 can be considered a working range and a pH of about 8.0 is preferred. Tris (hydrogen chloride / aqueous sodium chloride) is particularly suitable as a buffer.

The solubility of the Cg - C ^ q alkanecarboxylic acids is not high in water. Octanecarboxylic acid has a solubility of the order of 4 millimoles. Therefore the preferred procedure for the preparation of the eluent is that you first have a buffer, such as tris-hydrogen chloride (0.05-bis 0.2 molar) is expediently prepared with 0.05 to 0.5 molar aqueous sodium chloride and mixed with as much alkane

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carboxylic acid decomposes as it dissolves in it. The solution obtained must in turn have a pH of at least 6.5, with 7.5 being the working pH range and a pH of about 8.0 being preferred is. Regardless of the type of column regeneration, the columns can be used without loss of absorption power can be used repeatedly.

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Usable dyes

Color

Index names Trade name

17865 CI Reactive Cibacron Orange Orange G-E; Procion Brilliant Orange HGRS

61210 CI Reactive Cibacron

Blue 5 Brilliant Blue PBR-P

61211 CI Reactive Cibacron Blue Blue 2 P3G-A; Procion Blue HBS

Structural formula HO

-N = N-

SO ₃ Na

SO ₃ Na NaO ₃ S

Cl

ΪΨζ,

OjH

Cl / *

N N

SO

O ₃ H

SO ₃ H

NH-CO-NH,

18105 CI Reactive
Red 4

Cibacron
Brilliant Red 3B-A; Procion Brilliant Red H7BS

Usable dyes

Color index names Trade name

- -_

26440 CI Reactive Brown

Cibacron Brown 3GR-A; Procion Orange Brown HGS SO ₃ Na

Structural formula N

NH- f V-NH- (^ N SO, Na NN

17910 CI Reactive Red

CI Reactive Red

18156 CI Reactive Red

Cibacron Scarlet 2G; Procion Scarlet H3GS

Cibacron Scarlet 4G-P

Cibacron Brilliant Red B-A

CH ₃ CO

S0, well

⁵ HO
-N = N- f ^ r

NaO ₃ S ¹

S0, well

³ HO ..

Cl

SO ₃ Na

N =

Cl

NaOjSr ^^^ SO ₃ Na

18159 CI Reactive Red Cl

SO, Na HD // V-- NN -λ

νηΠνη-

HaO, ff "^^ eo ^ te

Usable dyes

Color

Index names Trade name

Blue dextran

Structural formula

O-dextran - NH-Q. * :: C. '.'. ^S0 3 ^H /

σ co oo

O NH ₂

O-dextran

Example I.

100 ml of settled crosslinked Sepharose '4B from Pharmacia Fine Chemicals, Inc. was suspended in 250 ml of water. 1 g of Cibacroir ^ Brilliant Blue FBR-P was dissolved in 25 ml of water and the solution was added to the Sepharose suspension. The mixture was stirred gently and ^{heated to 80 °} C. for 45 minutes. The pH was kept at at least 6 by adding sodium bicarbonate or tris-hydrogen chloride.The reaction mixture was cooled to room temperature, the supernatant liquid was decanted and the solid phase was washed thoroughly first with water (3 l), then with aqueous guanidine hydrochloride (5- molar _t 2 1) and finally suspended in a mixed buffer of aqueous tris-hydrogen chloride (0.05 molar) and aqueous sodium chloride (0.5 molar) at pH 8.0, whereupon a coupling product of Sepharose 4B and Cibacron Brilliant Blue FBR-P.

In the aforementioned method, settled Sepharose polyacrylamide gel and Amberlite ^ IRC 45 can be used to arrive at analogous coupling products.

According to the above procedure, but using Procion Brilliant Orange HGRS, Procion Blue HBS, Procion Brilliant Red H7B5, Procion Orange Brown HGS, Procion Scarlet H3GS, Cibacron Scarlet 4GP and Cibacron Brilliant Red BA as dyes and crosslinked Sepharose, polyacrylamide gel or a amino group-containing polyacrylic resin as a carrier phase obtained analogous coupling products, provided that they increased the reaction temperature to 90 to 95 ⁰ C.

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Example II
Manufacture of Sepharose / Dextran Blue Coupling Product

100 ml of settled Sepharose 4B from Pharmacia Fine Chemicals Inc. was reacted with an aqueous solution of 16 g of cyanogen bromide for five hours at a pH of 11.0 and a temperature of 10 ^° C. After the reaction had ended, the supernatant liquid passed through Removed decanting and stirred the Sepharose with 0.1 molar sodium bicarbonate buffer of pH 9.5. The supernatant liquid was again removed by decanting and 1 g of dextran blue in 100 ml of aqueous 0.1 molar sodium bicarbonate, pH 9.5, was added. The reactants were left to stand at 4 ^° C. for 24 hours. The supernatant liquid was then removed by decanting and the product was successively mixed with 4 liters of 0.1 molar aqueous sodium bicarbonate of pH 9.5, 4 liters of 6 molar aqueous urea, 4 liters of water and 4 liters of a mixed buffer of 0.05- molar aqueous tris hydrogen chloride and 0.5 molar aqueous sodium chloride of pH 8.0. After removing the supernatant washing liquid of the last-mentioned buffer, the coupling product obtained in this manner was suspended in a similar solution and stored at 4 ^0C.

Example III Removal of albumin from plasma

A chromatography column (1.0 χ 20 cm) was with about 10 ml of the coupling product prepared according to Example II, which is in the above-mentioned TrIs hydrogen chloride / sodium chloride buffer was suspended, loaded. 2 ml of plasma were added to the column and eluted with buffer until no further pro-

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no more was eluted. This was verified by examination of the effluent in the ultraviolet light which ^{A Weni} S ^he was ^{than 0 »020} 280th The eluate was then diaflowing using an Amicon-UM10 membrane, concentrated to a volume of 2 ml.

The process was controlled by taking comparative samples passed through a Sepharose 4B column which did not contain the coupling product with dextran blue contained. The diaflowed eluate was then eluted by cellulose acetate membrane electrophoresis analyzed, the results given in Table I below being obtained.

Segharο s e ^ Dex tran blue 0.16 % of
Control
Worth Table I. Control
plasma Hit the clutch
product from
Sepharose and
Dextran blue
treated
plasma 0.16 (Grams / 100 ml) 0.60 4th Obtaining plasma protein fractions according to Kontak 4.10 0.65 94 with 0.17 0.93 84 fraction 0.72 2.50 96 0.67 98 albumin 0.95 38 Alpha-1 Total protein 6.61 Alpha-2 beta gamma

The fractions were quantitatively analyzed after cellulose acetate membrane electrophoresis as described in the text. The total protein was determined by the method of Lowry et al (J. Biol. Chem. Vj £, 265 (1951).

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Example IV Removal of albumin from plasma

A chromatograph column (1.0 χ 20 cm) was charged with about 10 ml of the coupling product, which was prepared according to Example I using crosslinked Sepharose 4B as the carrier phase and Procion Brilliant Red H7BS as the dye, suspended in the above-mentioned iris (hydrogen chloride / sodium chloride) -Buffer solution prepared x 2 ml of plasma was added to the column and eluted with buffer until no more protein was eluted. This was verified by examining the eluate with ultraviolet light, whereby A ₂₈ Q was less than 0.020. The above-mentioned Tris sodium chloride buffer is then obtained as the eluate.

Example V Column regeneration without extracting albumin

The column according to Example IV was regenerated by running two column volumes of 6 molar aqueous urea through the column, followed by the Trls sodium chloride buffer mentioned above. This column regeneration process denatures the albumin, although it is perfectly satisfactory for the recovery of the column.

Example VI Regeneration of the column with albumin recovery

The albumin-containing column according to Example III was washed with two column volumes of 0.075 molar aqueous sodium phosphate of pH 2.4 and ethanol, with the ratio of ethanol to aqueous sodium phosphate was 1: 1. The eluate was then flowed using an Anicon UM-10 membrane obtained pure, undenatured albumin in an amount of 65 mg (about 80% recovery) became. 50 9811/106 2

Example VII - ■ - '

Re Generier ung ^ of säu le unte r Mbo Mngewin voltage

The column containing albumin according to Example III is washed with two column volumes of a mixture of 4 ° -molar octanoic acid solution, 0.05 molar Tris hydrogen chloride and O ₉ 05-smolare® aqueous sodium chloride at pH 8 ", the eluate maräe lyophilized, after which r® ± n @ 8> stabilized albumin was obtained.

In the case of the procedure mentioned, instead of ■■ ■ Qetanoic acid © nonane «· and decanoic acid can be used.

g JL A Jj,

Re ^ e nerierting the _{column r} under _i Mb uml ag e winnung

The albumin-containing column according to Example IV was washed with two columns of a mixture that was O ₉ O5 "ra © lar ®n calcium chloride and 0.05-raolar aa trie hydrogen chloride at pH 8.0" The eluate was dana d © »Diaflowing using a 1D» lfeabFan »- after which pure, not obtained *

IMrä® the CimlelomchloFid by lüagneaiumchlorid

ere © tet _p one obtained the same

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Claims (1)

Claims ·; Compound of the general formula 0 NH JAgaroseJ- O-dextran wherein R ¹ = H, R ² SO ₃ H or R ¹ - SO ₃ H, R * = H and χ is a weight ratio of 0.8 to 40. 2. A compound according to claim 1, characterized in that χ is 4 to 20 · 3. Compound according to claim 1, characterized, that agarose is in the physical form of Sepharose is present. 4 method for removing albumin from essentially cell-free aqueous fluids, marked by a) Suspending a ^ the general Formula Q3? | - JCRlJJ, where CRD is a reactive Dye of the general formula R ₁ O ₃ S -L 5.0 9 8 Γ1 / 1 0 6 2 i. NH η in the R _{3 is} an alkyl, phenyl, alkoxyphenyl, alkylphenyl rest, a radical of the formula J ⁼ \ - ^ 3 ¹¹ I or water AW / Substance means, where R _{1 is} hydrogen, an alkali metal or an alkaline earth metal and R _{2 is} halogen or -0-dextran Z ₁ J ~ -h] ₂ or -CH = CH-CH = CH-, Z _{2 is} a nitrogen-nitrogen bond or a hydrogen atom bonded to the nitrogen atom shown, Q denotes a substituted or unsubstituted aryl group when Z _{2 is} hydrogen, or denotes a substituted or unsubstituted aryl-N »group when Z _{2 is} a double bond, with the proviso that the substituent groups mentioned are not halogen groups or heavy metal groups, with under Heavy metal is understood to mean a metal that falls into groups of the periodic table other than groups 1 or 2A, and SP means a solid carrier phase consisting of agarose, polyacrylamide or acrylic resins containing aminogroups, and χ means a weight ratio between 0.8 and 40, in a buffer with a molarity of at least 0.05 at a pH of between 6.5 and 8.5, b) Inertihrung this suspension with the albumin-containing liquid to produce a Suspension with albumin adsorbed on the solid phase and c) Separation of the solid phase with the adsorbed albumin from the aqueous phase. 50981 1/1062 5. The method according to claim 4, characterized, that the coupling product is a substance of the formula used, wherein x, Q, Z ₁ , have the given meaning Z ₂ and that in claim 4 6. The method according to claim 5, characterized in that that a coupling product of the formula given in claim 5 is used, in which SP Sepharose, a polyacrylamide gel or an acrylic resin containing amino groups means. 7. The method according to claim 4 and 6, characterized in that that the coupling product is a substance of the general formula "O-dextran used, wherein χ and R _{1 have} the meaning given in claim 4. 5.O 9811/1062 8. The method according to claim 6, characterized in, that a coupling product of polyacrylamide gel and Procion Blue HBS, Cibacron Brilliant Blue FBR-P, Procion Orange Brown HGS, Procion Brilliant Red H7BS, Procion Scarlet H3GS or Procion Brilliant Orange HGRS used. 9. The method according to claim 4, characterized in that the solid phase with the albumin adsorbed thereon is also treated with an eluent consisting of aqueous urea, aqueous-ethanolic alkali metal phosphate, a base-buffered alkanecarboxylic acid with 2 to 10 carbon atoms or an aqueous solution of cations of group 2A of the periodic table and potassium as well as pharmaceutically acceptable anions and a basic buffer, treated with cleavage of the albumin from the solid phase. 10. The method according to claim 9 characterized in _t, that an alkanecarboxylic acid is used as the eluent 8 to 10 carbon atoms, which is buffered to a pH of 7.5 to 9, is used. 11. The method according to claim 10, characterized in that. that the alkanecarboxylic acid used is octanoic acid " 12. The method according to claim 9 _f, characterized in that that an aqueous solution of as the eluent Group 2A metal ions of the periodic table or potassium and pharmaceutically acceptable anions and a basic buffer is used, the metal ion has a 0.05 to 0.5 molar concentration. 50981 1/1062 13 · The method according to claim 9, characterized in that the albumin-free solid phase of the liquid phase separates, the albumin-free solid phase is reused in stage a) of claim 4 509811/1062