JPH06100592A - Composition, obtained by genetic manipulation and containing human serum albumin - Google Patents

Abstract

PURPOSE:To obtain the subject composition, containing a human serum albumin prepared by the genetic manipulation and no impurity components derived from the producing host, having high purity, capable of suppressing the coloring by coloring components and useful for treating, etc., hypoproteinemia. CONSTITUTION:The objective high-purity human serum albumin (HSA)- containing composition is obtained by transducing a recombinant plasmid containing a gene capable of coding the HSA integrated therein in a host cell such as Pichia yeast GTS115 strain (NRRL deposit No. Y-15851), carrying out the transformation, then culturing the resultant transformant in a culture medium at 30 deg.C temperature for 10-100hr and successively subjecting the resultant culture supernatant after completing the culture to the treatment with an ultrafiltration membrane, heat treatment and treatment with an acid, a cation exchanger, hydrophobic chromatographic carrier and an anion exchanger. This composition contains no impurity component derived from the producing host and has >=99.999999% purity of the human serum albumin.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a composition containing human serum albumin which is expressed by genetic engineering.

[0002]

2. Description of the Related Art Albumin, particularly human serum albumin (hereinafter also referred to as "HSA"), is a major protein constituent of plasma. This protein is made in the liver and is primarily responsible for maintaining normal osmotic pressure in the bloodstream. It also functions as a carrier for various serum molecules. HSA is administered in various clinical settings. For example, in patients with shock or burns, H2 is usually used to restore blood volume and thereby improve some of the trauma-related symptoms.
Requires frequent administration of SA. Patients suffering from hypoproteinemia or erythroblastosis may also require treatment with HSA. Therefore, the basic therapeutic significance of administering HSA lies in the treatment of conditions such as fluid loss from blood vessels, as in hypoproteinemia, which results in surgery, shock, burns, and edema. .

Currently, HSA is produced primarily as a product from a collected blood fraction. The drawbacks of this manufacturing method are that it is uneconomical and that the supply of blood is difficult. Blood may also contain undesired substances such as hepatitis virus. Therefore, HSA
It would be beneficial to develop alternative raw materials for.

With the advent of recombinant DNA technology, a variety of useful polypeptides can be produced by microorganisms, and many mammalian polypeptides have already been produced by various microorganisms. For HSA,
A technique for mass-producing by genetic engineering technology and highly purifying it is being established.

However, in genetic engineering, when culturing a microorganism as a host, and further when purifying HSA, some coloring components in the raw material or substances secreted by the microorganism become contaminated, which causes this. It is considered that HSA itself is colored by binding with HSA. Moreover, these contaminants are associated with conventional plasma-derived HSA.
It cannot be sufficiently removed by the above purification method.

[0006]

SUMMARY OF THE INVENTION Therefore, the object of the present invention is to obtain the above-mentioned coloring components and contaminants that could not be sufficiently removed by conventional methods for purifying plasma-derived HSA when obtaining HSA by genetic engineering. An object of the present invention is to provide an HSA-containing composition which is removed and whose coloring is sufficiently suppressed.

[0007]

Means for Solving the Problems As a result of intensive studies in view of the above circumstances, the inventors of the present invention have found that when obtaining HSA obtained by genetic engineering, particularly preferably at the final stage of the HSA purification step. It has been found that by treating the HSA with a chelating resin, a coloring substance is adsorbed to the chelating resin, the coloring of HSA can be reduced, and a composition containing high-purity HSA in which contaminants are sufficiently removed can be provided. The present invention has been completed.

[0008] That is, the present invention is a composition containing HSA obtained by genetic engineering and having the following characteristics: (1) Substantially free of contaminant components derived from the production host (2) HSA Purity is 99.999999% or higher.

The HSA-containing composition having the above-mentioned characteristics is a chelating resin for the HSA obtained by genetic engineering, particularly preferably at the end of the purification step.
Preferably, it is obtained by treating with a chelate resin having an exchange group having a chelating ability selected from a polyol group, a polyamine group or a thiourea group as a ligand.

The present invention provides H obtained by genetic engineering.
The present invention relates to a SA-containing composition, wherein the HSA is a bacterial cell that expresses HSA through genetic manipulation (for example, E. coli,
Yeast, Bacillus subtilis, koji, animal cells, etc.) are cultured and produced by extracellular expression (secretion expression).

(1) HSA Obtained by Genetic Manipulation The HSA-producing host prepared by genetic engineering used in the present invention is not particularly limited as long as it is prepared by genetic engineering, and it has already been described in known literature. In addition to the ones, those developed in the future can be appropriately used. Specific examples include bacteria (eg, Escherichia coli, yeast, Bacillus subtilis, etc.) that have been made HSA-producing through genetic manipulation, animal cells, and the like. In particular, in the present invention, as a host, yeast, especially Saccharomyces (for example, Saccharomyces cerevisiae)
], Or the genus Pichia [for example, Pichia pastoris (Pichia pastoris)] is preferably used. Also, auxotrophic strains and antibiotic sensitive strains can be used. Furthermore, Saccharomyces cerevisiae AH22 strain (a, his 4, leu 2, can 1), Pichia pastoris GT
S115 strain (his 4) and the like are preferably used.

Methods for preparing these HSA-producing hosts, methods for producing HSA by culturing the same, and HS from cultures
All the separation and collection methods of A are carried out by adopting publicly known methods and their modifications. For example, HSA
Examples of the method for preparing the producing host (or HSA producing strain) include a method using a normal human serum albumin gene (Japanese Patent Laid-Open Nos. 58-56684, 58-90515, 58-150517), and novel human serum. Method using albumin gene (JP-A-62-29985, JP-A-1-98486), method using synthetic signal sequence (JP-A-1-240191), method using serum albumin signal sequence (JP-A-2- 167095
JP-A-3-72889), a method of fusing hosts with each other (JP-A-3-53877), a method of causing mutation in a medium containing methanol, Method using mutant AOX ₂ promoter (Japanese Patent Application No. 3-63598, 3-
No. 63599), expression of HSA by Bacillus subtilis (Japanese Patent Laid-Open No. Sho 6-62 (1994)).
2-25133), expression of HSA by yeast (JP-A-60-41487, 63-39576, 63-63).
74493), the expression of HSA by Pichia yeast (JP-A-2-104290) and the like.

Of these, the method of causing mutation in a medium containing methanol is specifically performed as follows. That is, first a suitable host, preferably Pichia yeast, specifically GTS115 strain (NRRL Deposit No. Y-15851).
A transformant is obtained by introducing a plasmid having a transcriptional unit expressing HSA under the control of the AOX ₁ promoter into the AOX ₁ gene region of E. coli (JP-A-2-104).
290). This transformant has weak growth ability in methanol medium. Therefore, this transformant is cultured in a medium containing methanol to cause a mutation, and only a viable strain is recovered. At this time, the methanol concentration is, for example, about 0.0001 to 5%. The medium may be an artificial medium or a natural medium. Culture condition is 1
The temperature is, for example, 5 to 40 ° C. and 1 to 1000 hours.

As a method for culturing an HSA-producing host (that is, a method for producing HSA), in addition to the methods described in the above-mentioned publications, a high concentration of glucose can be supplied in appropriate small amounts by fed-batch culture. Then, a method of obtaining a high-concentration bacterial cell and a product by avoiding high-concentration substrate inhibition on the producing bacterial cell (Japanese Patent Application No. 1-219561), and a method of adding a fatty acid to the medium to enhance HSA production ( Japanese Patent Application No. 3-81
No. 719) is exemplified. Further, as a method for separating and collecting HSA, in addition to the methods described in the above-mentioned respective publications, inactivation of protease by heat treatment (Japanese Patent Laid-Open No. 1031/1991).
88), an anion exchanger, a hydrophobic carrier, and at least one selected from the group consisting of activated carbon, and HSA is used.
And a coloring suppression method by separating the coloring component (Japanese Patent Application Laid-Open No. 4-54198).

The medium used for culturing the transformed host is
Usually, a medium known in the art to which a fatty acid having 10 to 26 carbon atoms or a salt thereof is added is used, and the culture condition is carried out according to a general conventional method. The medium may be a synthetic medium or a natural medium, and a liquid medium is preferable. For example,
As a synthetic medium, generally, various sugars as carbon sources, urea, ammonium salts, nitrates and the like as nitrogen sources, various vitamins and nucleotides as micronutrients, and Mg, Ca, Fe, Na, K, Mn and Co as inorganic salts are generally used. , Cu
Are exemplified. YNB liquid medium [0.7% yeast nitrite base (manufactured by Difco), 2% glucose] and the like can be mentioned. As a natural medium, YPD
A liquid medium [1% yeast extract (manufactured by Difco), 2% bactopeptone (manufactured by Difco), 2% glucose] is exemplified. The pH of the medium may be neutral, weakly basic or weakly acidic. Further, in the case of a methanol-assimilating host, a methanol-containing medium can be used. In this case, the methanol concentration is about 0.01-5%.

The culturing temperature is 15 to 43 ° C (20 to 20 ° C for yeast).
30 ° C., and bacteria preferably 20 to 37 ° C.). The culturing time is about 1 to 1000 hours, and the culturing is allowed to stand or shake,
It is carried out by a batch culture method, a semi-batch culture method or a continuous culture method under stirring and aeration. In addition, it is preferable to perform pre-culture prior to the culture. As the medium at this time, for example, YNB liquid medium or YPD liquid medium is used. The culture conditions of the preculture are as follows. That is, the culture time is preferably 10 to 100 hours, and the temperature is preferably about 30 ° C. for yeast and about 37 ° C. for bacteria.

After the completion of the culture, HSA is collected from the culture filtrate, the cells or the cells by known separation means.

(2) Purification of HSA As the purification step of HSA of the present invention, known methods such as various fractionation methods, adsorption chromatography, affinity chromatography, gel filtration, density gradient centrifugation, dialysis and the like are adopted. It Suitable examples of the purification step include the following steps including: The culture supernatant of the human serum albumin producing host is treated with ultrafiltration membranes having a molecular weight cutoff of 100,000 to 500,000 and 1,000 to 50,000. Heat treatment is performed at 50 to 70 ° C. for 30 minutes to 5 hours. Acid treatment at pH 3-5. Treatment is carried out using an ultrafiltration membrane having a molecular weight cutoff of 100,000 to 500,000. After contacting with a cation exchanger under conditions of pH 3 to 5 and salt concentration of 0.01 to 0.2M, elution is performed under conditions of pH 8 to 10 and salt concentration of 0.2 to 0.5M. The non-adsorbed fraction is recovered by contacting the carrier for hydrophobic chromatography under the conditions of pH 6 to 8 and salt concentration of 0.01 to 0.5M, and pH 6 to 8 and salt concentration of 0.01 to 0.1M. The non-adsorbed fraction is collected by contacting the anion exchanger under the conditions. Further, instead of the above steps, pH 6 to 8 and salt concentration 1 to
After contacting the carrier for hydrophobic chromatography under the condition of 3 M, elution under the conditions of pH 6 to 8 and salt concentration of 0.01 to 0.5 M, or instead of the above process, pH 6 to
8. After contacting with an anion exchanger under the condition of salt concentration 0.001-0.05M, pH 6-8, salt concentration 0.05
The step of eluting under the condition of ~ 1M, and further between, or after the step, salting out under the conditions of pH 3-5 and salt concentration 0.5-3M, and the precipitation fraction. May be further included.

(3) Decolorization of HSA The decolorization step of HSA of the present invention comprises the steps of:
Particularly preferably, it is carried out by bringing HSA into contact with a chelating resin which is incorporated at the end and has a specific ligand portion. The carrier portion of the chelate resin is preferably a carrier having hydrophobicity, and examples thereof include a copolymer of styrene and divinylbenzene and a copolymer of acrylic acid and methacrylic acid. On the other hand, the ligand part is a polyamine group having a plurality of polyol groups such as N-methylglucamine group, imino group, amino group, ethyleneimino group, etc. in the molecule (including polyalkylene polyamine groups such as polyethylene polyamine). Included), and thiourea groups. Commercially available chelate resins having the above-mentioned carrier portion and ligand portion include DIAION CRB02 (ligand portion; N-methylglucamine group, manufactured by Mitsubishi Kasei), in which the carrier portion is a copolymer of styrene and divinylbenzene.
CR20 (ligand _{moiety; -NH (CH 2 CH 2 NH} ) n H, manufactured by Mitsubishi Kasei), LEWATIT TP (ligand portion; -NHCSN
H ₂ manufactured by Bayer) and Amberlite CG4000 are preferably used.

The treatment conditions with the chelate resin are preferably as follows. pH condition: acidic or neutral (3-9, preferably 4-)
7) Time: at least 1 hour or more, preferably 6 hours or more Ionic strength: 50 mmho or less, preferably 1 to 10 m
mho mixing ratio: 0.1 to 100 of resin for 250 mg of HSA
g, preferably 1-10 g (wet weight)

The degree of coloring of HSA obtained through the above-mentioned steps (including treatment with salting out and further treatment with chelating resin) is A _{500 nm} / A _{280 nm in} the case of 25% HSA solution.
Is about 0.001 to 0.005. The chelating resin treatment of the present invention reduces the coloring degree of HSA to 1/2 to 1/10. In particular, the absorption degree near 500 nm, that is, the reddish coloring degree is reduced to 1/3 to 1/10.

(4) Formulation The obtained HSA (containing composition) can be formulated by a known method (ultrafiltration, addition of a stabilizer, sterilization filtration, dispensing, lyophilization, etc.). . HS prepared in this way
The preparation A can be clinically used as an injection similar to the plasma-derived HSA preparation. It can also be used as a stabilizer, carrier or carrier for pharmaceuticals. The HSA-containing composition means that the HSA of the present invention has a high purity but is not a pure product, and some impurities are mixed even in a trace amount. However, the state in which additives (for example, biologically acceptable carrier), excipients, stabilizers, etc. are added is excluded.

(5) Purified genetically engineered HSA
Properties of (Containing Composition) The HSA of the present invention has a molecular weight of about 67,000 and an isoelectric point of 4.6 to.
It is a single substance of 5.0. The HSA-containing composition is composed of only a monomer and does not substantially contain a dimer, a polymer or a decomposed product. Specifically, the total content of dimers, polymers and decomposed products is about 0.01% or less. Further, it is substantially free of contaminant components (eg, proteins, polysaccharide components, etc.) derived from the production host. Specifically, in the case of a 25% HSA solution, the protein component is 1 ng / ml or less, preferably 0.1 ng / ml or less. Similarly, the polysaccharide component is 10 ng / ml or less, preferably 1 ng / ml.
The following is exemplified. As a result, the purity of HSA is 9
9.999999% or more, preferably 99.99999
99% or more is exemplified. Coloring degree is A ₃₅₀ / A ₂₈₀ in the case of the HSA 25% solution 0.01 to 0.05, A ₄₅₀
/ A ₂₈₀ is 0.001~0.02, A ₅₀₀ / A ₂₈₀ is illustrated about .001 to .005. Also, H
The amount of fatty acid bound to SA is 1 molecule or less, preferably 0.1 molecule or less per 1 molecule of HSA.

[0024]

INDUSTRIAL APPLICABILITY According to the present invention, with respect to HSA obtained by genetic engineering, certain coloring components in raw materials or substances secreted by microorganisms are contaminated, and coloring caused by binding with HSA is sufficient. It is possible to provide a composition containing high-purity HSA in which impurities are sufficiently suppressed and which is sufficiently suppressed.

[0025]

EXAMPLES In order to explain the present invention in more detail, examples will be given, but the present invention is not limited thereto.

Reference Example 1 Cultivation of HSA-producing host (1) Strain used: Pichia pastoris GCP101 strain Pichia pastoris GTS11 was prepared by the method described in JP-A-2-104290.
The AOX1 gene region of 5 (his4) was replaced with the fragment cut with NotI of the plasmid pPGP1 having a transcription unit expressing HSA under the control of the AOX1 promoter to obtain PC4130. This strain is AOX1
Due to the absence of the gene, the ability to grow in a medium containing methanol as a carbon source is low (Mut- strain).

PC4130 was inoculated into 3 ml of YPD medium (1% yeast extract, 2% bactopeptone, 2% glucose), and after 24 hours, initial OD ₅₄₀ = 0.1.
To 50 ml of YPD medium. 3 days 30
YPD so that the initial OD ₅₄₀ = 0.1 after culturing at ℃
50 ml of medium was inoculated. The same passage was repeated every 3 days. The cells were diluted with sterilized water so that the number of cells was 10 ⁷ cells / plate at each passage, and 2% MeOH-YNBw / oa.
a. Plate (0.7% yeastite rosin ene base with out amino acid, 2% methanol, 1.5
% Agar powder was applied and cultured at 30 ° C. for 5 days to determine the presence or absence of colonies. As a result, 2 applied after 12 days passage
% MeOH-YNBw / oa. a. The plate yielded 20 colonies. Mut-strains can hardly grow on this plate, while Mut + strains can grow. That is, the fact that colonies were formed on this plate indicates that the assimilation ability of methanol was increased and a strain converted to Mut + was obtained. One of the resulting colonies was appropriately diluted with sterile water and diluted with 2% MeOH-YNBw / o.
a. a. It was spread on a plate and isolated into a single colony.
One of them was named GCP101.

(2) Culture of strain (pre-preculture) 20 ml of 1 ml of glycerol frozen stock strain
Baffled 1,000 with 0 ml YPD medium (Table 1)
The cells were inoculated into an Erlenmeyer flask of ml volume and cultured at 30 ° C. for 24 hours with shaking.

[0029]

[Table 1]

(Pre-culture) A pre-preculture medium was inoculated into a 10-L jar fermenter containing 5 L of YPD medium, and agitated and cultured for 24 hours. Culture temperature is 30 ℃, aeration rate is 5L /
Minutes Further, the pH was not controlled in the preculture.

(Main culture) Medium for batch culture (Table 2) 25
The preculture liquid was inoculated into 0 L and cultured with aeration and stirring using a 1,200 L volume fermenter. The tank pressure is 0.5 kg / c
Batch culture was started while controlling the stirring speed so that the dissolved oxygen concentration was maintained at about 50% to 30% of the saturated dissolved oxygen concentration with m ² and the maximum aeration amount set to 800 NL / min.
The addition of the feed medium (Table 3) was started when the glycerol in the medium was consumed in the batch culture. A computer was used to add this feed medium, and high-density culture was performed while controlling so that methanol did not accumulate in the medium. The pH was controlled to a fixed value of pH 5.85 by adding 28% ammonia water. For defoaming, add an antifoaming agent (Adecanol, manufactured by Asahi Denka Kogyo Co., Ltd.) to 0.3
It was carried out by adding 0 ml / L in advance and then adding a small amount as needed.

[0032]

[Table 2]

[0033]

[Table 3]

Reference Example 2 AOX2 promoter [mutant type] isolated from the GCP101 strain of Reference Example 1. Natural AOX2 promoter (TEAST,
5, 167-177 (1988) or Mol. Cell, Biol., 9, 1316-1
323 (1989)), the HSA expression plasmid pMM042 was constructed by mutating the 255th base upstream of the start codon from T to C].
pastoris) introduced into GTS115 strain and transformed into UHG
42-3 strain was obtained (Japanese Patent Application No. 3-63599). This UHG42-3 strain was cultured according to Reference Example 1 to produce HSA.

Reference Example 3 Purification of HSA [i] Separation of culture supernatant to membrane fractionation (II) About 800 L of the culture solution obtained in Reference Example 1 was pressed to separate the culture supernatant. The culture supernatant has a molecular weight cutoff of 30.
Treated with 10,000 ultrafiltration membranes. Then, the liquid amount was concentrated to about 80 L using an ultrafiltration membrane having a molecular weight cut off of 30,000 [membrane fraction (I)]. This concentrated solution was heat-treated at 60 ° C. for 3 hours, then rapidly cooled to about 15 ° C., adjusted to pH 4.5, and again treated with an ultrafiltration membrane having a molecular weight cutoff of 300,000. Picture (II)]. Then, using an ultrafiltration membrane with a cut-off molecular weight of 30,000, the buffer solution in the alummine solution was adjusted to 50 mM.
50 mM acetate buffer containing sodium chloride, pH 4.5
I replaced it with.

[Ii] Cation exchanger treatment 50 mM acetate buffer containing 50 mM sodium chloride, p
After adsorbing albumin on an S-sepharose packed column equilibrated with H4.5 and thoroughly washing with the same buffer,
0.1M phosphate buffer containing 0.3M sodium chloride,
Elution of albumin was performed at pH 9.

[Iii] Hydrophobic Chromatography Treatment The albumin solution eluted from the column packed with S-Sepharose was loaded onto a column packed with phenylcellulofine equilibrated with 50 mM phosphate buffer containing 0.15 M sodium chloride, pH 6.8. Was added. Under these conditions, the fraction that passed through the column was collected without adsorbing phenylcellulofine. The albumin that passed through the column was concentrated to a volume of about 50 L using an ultrafiltration membrane having a molecular weight cut off of 30,000, and the buffer solution in the albumin solution was replaced with a 50 mM phosphate buffer solution, pH 6.8.

[Iv] Anion exchanger treatment After the hydrophobic chromatography treatment, the concentrated and buffer-exchanged albumin solution was added to a column packed with DEAE-Sepharose equilibrated with 50 mM phosphate buffer, pH 6.8. . Under these conditions, albumin passed through the column without being adsorbed on DEAE-Sepharose.

[V] HSA salting-out treatment A solution having a final concentration of 1 M by adding sodium chloride to 5% concentration of HSA was adjusted to pH 3.5 with acetic acid to prepare HSA.
Was allowed to settle. This precipitate was separated from the supernatant by centrifugation to remove impurities.

[Vi] Decolorization treatment 1 ml of purified HSA having a concentration of 25% was added to DIAION CRB.
02 (carrier part is styrene-divinylbenzene copolymer, ligand part is chelating resin consisting of N-methylglucamine group, manufactured by Mitsubishi Kasei), 1 g is added, and pH is 6.8, ionic strength is 5 mmho, and room temperature is for 24 hours. It was stirred. The resin was washed with distilled water and the non-adsorbed fraction was recovered as a HSA-containing composition.

Example 1 Properties of Purified HSA (Composition Containing Composition) of the Present Invention (1) HPLC Analysis of Purification Step HSA after the purification step up to hydrophobic chromatography was analyzed by HPLC gel filtration. Gel filtration analysis was performed under the following conditions. Column: TSK gel G3000SW (manufactured by Tosoh Corporation) Developer: 0.1M KH ₂ PO ₄ /0.3M NaCl buffer Detection: Absorbance at wavelength 280 nm The purified HSA-containing composition became a single peak of HSA monomer.

(2) Yeast-Derived Component Analysis A culture supernatant of albumin non-producing yeast was roughly purified by the same method as this method to immunize a rabbit, and the obtained antiserum was used to prepare a purified HSA-containing composition. The yeast-derived components present in the plant were detected. The measurement was performed by enzyme immunoassay (EIA method). The detection result of the yeast-derived component for each sample is shown. The sample was measured using an albumin concentration adjusted to 250 mg / ml.

[0043]

[Table 4]

(3) Molecular weight The molecular weight was measured by the above-mentioned HPLC gel filtration method. The molecular weight of HSA in the purified HSA-containing composition of the present invention is about 67.
000, which was comparable to plasma-derived HSA.

(4) Isoelectric point The isoelectric point was measured using a thin layer polyacrylamide gel according to the method of Allen et al. [J. Chromatog., 146, 1 (1978)]. The isoelectric point of HSA in the purified HSA-containing composition of the present invention was about 4.9, which was similar to that of plasma-derived HSA.

(5) Coloring degree The coloring degree is 280 nm, 350 nm, 450 nm, 500
Absorbance at nm is measured, A ₃₅₀ / A ₂₈₀ , A ₄₅₀ / A
₂₈₀ and A ₅₀₀ / A ₂₈₀ were calculated. Purified HSA of the present invention
The coloring degree of the contained composition is A ₃₅₀ / A ₂₈₀ is about 0.02, A
₄₅₀ / A ₂₈₀ is about 0.01, A ₅₀₀ / A ₂₈₀ is about 0.0
02, which was similar to that of plasma-derived HSA.

(6) NFEA-Test Wako (manufactured by Wako Pure Chemical Industries) was used for measuring the amount of bound fatty acid. As a result, it was 1.6 mol (per 1 mol of HSA) before the chelate resin treatment, but 0.03 after the treatment.
It significantly decreased to 7 mol (per mol of HSA).

Example 2 The culture solution obtained in Reference Example 2 was treated in the same manner as in Reference Example 3. The properties of the purified HSA (containing composition) were similar to the molecular weight, isoelectric point and coloring degree, polysaccharide content, gel filtration pattern, and yeast-derived component amount disclosed in Example 1.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical display location (C12P 21/02 C12R 1:84) (C12P 21/02 C12R 1: 865) (72) Inventor Kaideko Uede 2-25-1 Otani Otani, Hirakata-shi, Osaka Prefecture Midori Cross Central Research Institute Co., Ltd. (72) Inventor Munehiro Noda 2-25-1 Otani Otani, Hirakata City, Osaka Pref. (72) Inventor Takao Omura 2-25-1 Otani Otani, Hirakata City, Osaka Prefecture Midori Central Research Institute Co., Ltd. (72) Kazumasa Yokoyama 2-25-1 Otani Invitation, Hirakata City, Osaka Prefecture Midori Corporation Cross Central Research Institute

Claims (1)

[Claims] 1. A human serum albumin-containing composition containing human serum albumin obtained by genetic engineering and having the following characteristics: (1) Substantially free of contaminant components derived from a production host (2) Human serum Purity of albumin is 99.999999%
that's all.