JP2003501480A - Production method and preparation of intravenous immunoglobulin - Google Patents

Abstract

  (57) [Summary] The virus inactivation by the heat treatment of the gamma globulin solution is substantially free of virus, and the gamma globulin is further subjected to virus inactivation with a solvent-surfactant without precipitating the purified desired gamma globulin. A continuous production method for producing an intravenously injectable gamma globulin solution, characterized by the following: Partially purified gamma globulin solids are not recovered as intermediates during the disclosed process.

Description

Detailed Description of the Invention

BACKGROUND OF THE INVENTION The present invention relates to a multi-step industrial process which is essential for the production of intravenously injectable immunoglobulins containing IgG (γ-globulin) as a main component.

Various steps for obtaining an intravenously injectable γ-globulin solution using a corn fraction of human plasma as a starting material are known. Some corn fractions contain higher titers of γ-globulin than others. The usual starting material for the γ-globulin solution is Cohn fraction II or Cohn fraction II + III.

The prior art employs various separation and sterilization techniques, and process changes have always been made with the end product purity and safety in mind, and overall yield improvements.

In many industrial processes, the solvent / surfactant process is the virus inactivation process,
Alternatively, one of the heat treatment steps is adopted as the virus inactivation step. To date, in the prior art, a corn fraction II paste or II + III paste was used as a starting material, and a multi-step treatment process including two different virus inactivation treatment methods effective in each part was used to obtain a continuous high yield γ. -No globulin production process was provided.

US Pat. No. 5,151,499 to Kameyama et al. Uses solvent / detergent treatment as a method of virus inactivation of coated viruses in protein compositions and heat treatment as a method of virus inactivation of membraneless viruses in protein compositions. The present invention is directed to viral inactivation treatment of protein compositions. The '499 patent teaches that solvent / detergent treatment is preferably performed first in the presence of a protease inhibitor, followed by heat treatment. The heat treatment is carried out in the liquid state and before any heat treatment the protein is first recovered from the solvent / surfactant by adsorption on the ion exchange column.
The liquid heat treatment is performed in the presence of a stabilizer which is sugar, sugar alcohol or amino acid. The '499 patent describes a number of starting protein compositions that include immunoglobulins, but employs Factor IX, thrombin, fibrinogen and fibronectin as manufacturing examples. The removal of denatured proteins in the heat treatment step during fractionation was not considered.

US Pat. No. 5,371,196 to Yuki et al. Is directed to the purification of secretory immunoglobulin A. Inactivation by liquid heat treatment or a combination of various liquid heat treatments and solvent treatments is described. The polyethylene glycol fraction is employed following each step and in the final step. This patent does not relate to high titers of γ-globulin.

One prior art process for the process of intravenous γ-globulin solutions is the corn fraction
Liquid heat treatment in the presence of sorbitol in a multi-step purification process using II + III paste as a starting material will be described. Hirao et al., U.S. Pat.
EG followed by 12% w / v PEG) and ion exchange chromatography (
Human blood group antibody is removed by DEAE-Sephadex, sorbitol is added as a protein stabilizer, and liquid heat treatment is performed. On the other hand, in Example 1 of US Pat. No. 4,876,088 to Hirao et al., The paste was suspended in water, the pH was adjusted to 5.5 and centrifuged, and the supernatant was treated with 33% w / v sorbitol for virus inactivation. And heat-treated under PEG fractionation (6
% / 12%) and other purification treatments including DEAE-Sephadex ion exchange chromatography are described, and a γ-globulin solution from an injectable Cohn Fraction II + III paste is described.

An object of the Summary of the Invention The invention is to provide an industrial process essential multistage for the production of highly purified γ- globulin solution by Cohn fractionation method.

Another object of the invention is to provide a highly pure injectable γ-globulin solution free of both capsular and membraneless viruses, including heat sensitive viruses.

A further object of the present invention is to provide for removal of any denatured proteins produced during heat treatment in an industrial γ-globulin process prior to the second virus inactivation treatment.

A further object of the present invention is to provide continuous industrial gamma-globulin without the need for intermediate gamma-globulin protein recovery for heat sterilization, PEG fractionation, and solvent detergent virus inactivation. Is to provide the production of.

The alcoholic fraction, which may be partially purified but is not rich in γ-globulin, is heat treated in the presence of a heat stabilizer in an aqueous medium for virus inactivation, The heat-treated solution is then first PEGylated and then in the presence of a solvent, preferably in the presence of a solvent-surfactant mixture, without the intermediate γ-globulin protein being recovered, in order to disrupt the coat virus. The above and other objects will be apparent to those skilled in the art in the light of the present invention provided with a second viral inactivation treatment followed by removal of the solvent or solvent-detergent mixture.

In one preferred embodiment of the invention, the collected PEG fractionation products are contaminated with bentonite for further virus removal prior to virus inactivation treatment with a solvent, preferably solvent-detergent. .

In a preferred embodiment of the present invention, sorbitol is the heat stabilizer and trialkyl phosphate is the solvent.

In another preferred embodiment of the present invention, the denaturation product by the heat-induced virus inactivation treatment is removed by the PEG fractionation before the second virus inactivation treatment, so that heat-treated γ-globulin having an extremely high purity is provided. It

In another preferred embodiment of the invention, any particulates present are removed prior to the solvent-surfactant treatment.

In a preferred embodiment of the invention, the γ-globulin solution is treated with an anion exchange resin.

In a preferred embodiment of the present invention, the one-step polyethylene glycol fractionation step comprises
It is carried out without precipitating the desired γ-globulin.

In a further preferred embodiment of the present invention, the γ-globulin solution is treated with a cation exchange resin, or diafiltration and / or tangential flow filtration.
The virus inactivation is completed.

A still further preferred embodiment of the present invention comprises the provided heat-sterilization treatment,
The solvent-surfactant-sterilized γ-globulin is suitable for intravenous injection.

The manufacturing method disclosed herein DETAILED DESCRIPTION OF THE INVENTION is a continuous process, the impure starting fraction containing immunoglobulin like Cohn fraction II + III paste, was once process starts Then, the process is carried out without removal of intermediate, partially purified gamma globulin material until it is complete (until highly purified gamma globulin can be supplied as a result product). Notably, the partially purified gamma globulin paste was not recovered as an intermediate product during the polyethylene glycol fractionation steps disclosed herein.

Fractions containing immunoglobulins are used as starting material. This fraction is of human origin and is not particularly limited as long as it is a fraction containing immunoglobulin. In a definite example, the paste containing the fraction II + III-containing and the fraction II-containing immunoglobulin-containing fraction and the immunoglobulin-containing fraction paste obtained in the ethanol fraction of Cohn is equivalent. Other starting materials include Fraction I + II + III, and Fraction II + IIIw and Fraction III paste. The starting material may include human blood-type antibodies, plasminogen, plasmin, kallikrein, prekallikrein activator, IgM, IgA, IgG polymer (hereinafter "aggregate"), and other impurities.

The preferred starting material is corn fraction II + III or corn fraction II paste. The Cohn Fraction II + III can be used as the starting material, or first a preliminary wash to form Fraction II + IIIw can be performed, which is then used in the process of the invention. "Fraction II + IIIw" is the corn fraction washed with disodium phosphate solution
It is a precipitate of II + III.

Fraction II + IIIw contains 20% of fraction II + III precipitate per about 1 kilogram of II + III paste.
Obtained by suspending in double volume of cold water for injection. Sodium phosphate solution is added to dissolve lipids, lipid proteins and albumin to a final sodium phosphate concentration of about 0.003M. Cold ethanol is added to give a final alcohol concentration of about 20%. During the addition of alcohol, the temperature is gradually lowered to -5 ± 1 ° C and the pH is adjusted to 7.2 ± 0 with, for example, acetate buffer or diluted hydrochloric acid.
Maintained at .1 and adjusted. Fraction II + IIIw precipitate is obtained by centrifugation and / or filtration while maintaining the temperature at -5 ± 1 ° C.

In the first virus inactivation step of the present invention, various preliminary purification and / or aggregate-reduction treatments are performed. For example, if Fraction II + IIIw paste was used,
Generally, about 20% alcohol and 70% or more protein are contained as IgG, which has a temperature of 0 to 5 ° C. and has a pH value using acetate buffer or diluted hydrochloric acid.
It is suspended in cold water adjusted to 4.5 to 6.0, preferably 5.0 to 5.5, in a volume of 3 to 20 times, preferably 10 to 15 times. The mixture is stirred for 2 to 15 hours so that all γ-globulin goes into solution. Then albumin and α-
Insoluble proteins such as globulins are removed by centrifugation and / or filtration.

When starting with the Fraction II + III paste, each kilogram of Fraction II + III paste is suspended in 3 to 20 kg, preferably 13 to 17 kg of cold water. The pH of the suspension is adjusted to 4.5 to 6.0, preferably 4.8 to 5.4. Suspension from 1 to 2
Mix for 0 hours, preferably 2 to 10 hours, at 0 to 25 ° C, preferably 0 to 10 ° C. Insoluble material is removed by filtration or centrifugation with a depth membrane. If filtration is performed, about 1 to 5 w / w% filter aid is used prior to separation. The centrifuge or filtrate is further clarified by a membrane membrane. The clarified solution is ultrafiltered through a 10,000 to 100,000 molecular weight cut-off (MWCO) membrane and concentrated to a protein concentration of about 1 to 12%, preferably 4 to 8%.

If different Cohn fractions are adopted for the starting, in the first step or step of the step, in order to obtain a fraction containing high IgG in a further step, the desired preliminary purification, selected appropriately, It can be performed. For example, Cone Fraction II (95% or more Ig
(Including G) is separated from Cohn Fraction III, Fraction II is further processed,
The first treatment, as described in Uemura et al., US Pat. No. 4,371,520, is an immunoglobulin aggregate contaminated in immunoglobulin monomers and dimers, which aggregate is known to cause anti-complement activity (ACA). , To remove the pH
Acid treatment from 3.2 to 5.0, preferably 3.8 to 4.2. Alternatively, if corn fraction II + III was the starting material, the low pH treatment of the Uemura et al. Patent followed the first purification step, followed by a heat treatment virus inactivation step as described above. , As an additional step.

In the heat sterilization step, the liquid mixture of immunoglobulin proteins is a partially purified product as described above, such as a filtrate from a purified Fraction II + III paste.
Concentration by ultrafiltration to about 1 to 12%, preferably 4 to 8% and addition of sugars, sugar alcohols and / or amino acids as heat treatment stabilizers is performed.
The heat stabilizer is preferably sucrose, maltose, sorbitol or mannitol, most preferably sorbitol. The sugar or sugar alcohol is used as an immunoglobulin solution in the form of powder or first mixed with a small amount of water, and the final concentration is about 25 to 50 w / w% to be saturated, preferably 30 to 40 w / w%. Is. In this respect, an aqueous solution of immunoglobulins contains sufficient water, therefore it contains about 1 to 8% total protein, and the typical starting material Fraction II + III is 1 300 per kilogram of paste
g protein.

Following the addition of the heat treatment stabilizer, the pH of the solution is adjusted to 4.5 to 6.5, preferably 5.0 to 6.0, and the mixture is adjusted to 50-7 for the inactivation of the heat-sensitive virus.
Heat at 0 ° C for about 1-20 hours, preferably 10-11 hours at about 60 ° C. The heat treatment step not only inactivates the virus, but also has the effect of preferentially denaturing undesired proteins such as prekallikrein, plasmin, plasminogen and IgA related to Cohn fraction II + III.

After the heat treatment, the solution is diluted directly or with cold water up to 5 times the amount of the heat treatment. The solution is then cooled to 0-10 ° C, preferably 0 to 5 ° C.

Next, the PEG fraction is applied to the heat treatment solution. PEG fractionation is a well known technique for separating desired IgG monomers and dimers from IgG aggregates and other naturally occurring impurities in the starting plasma protein fraction in immunoglobulin purification processes. Processing. However, in the current method, the PEG fractionation method also achieves the separation of desired IgG monomers and dimers from undesired denatured proteins produced by heat treatment. These denatured protein products include denatured prekallikrein activator, plasminogen, plasmin, IgA, I
gM and aggregates.

PEG concentration in the PEG fractionation step described in the prior art is to precipitate undesired proteins such as aggregates and eliminate them from solution, leaving the desired IgG monomers and dimers in solution. Any conditions of pH and pH can be selected. The PEG is provided as a powder, flakes or a 50% solution and is directly supplied to the heat-treated solution at a desired PEG concentration.

For example, when the PEG fraction is the starting material of Fraction II + IIIw paste,
A pH of between about 5.0 and 7.5, preferably between about 6.0 and 7.5, and, when the Fraction II + III paste is the starting material, preferably between about 5.5 and 6.0 and a PEG concentration of about 4;
To 8%, preferably 4 to 6% when the fraction II + IIIw paste is the starting material, and 6 to 8% when the fraction II + III paste is the starting material. You can While maintaining a cold temperature of about 0-2 ° C, PEG fractionation is carried out for about 1 to 8 hours, after which the precipitate is removed by either centrifugation or filtration.

In the optional virus removal step, bentonite is added to a final concentration of about 0.05 to 2.0 w / w%, preferably 0.1 to 1.0 w / w% for the centrate or filtrate, and the mixture is 1 For 5 hours and the bentonite paste is removed by filtration.

The final essential step of the invention is that the second virus inactivation treatment uses a solvent or solvent-detergent mixture. As will be described below, in particular ion exchange resins can be used as a further purification step before and / or after the solvent-surfactant treatment. An optional treatment is an anion exchange treatment to further remove albumin, transferrin and prekallikrein activators prior to the solvent detergent virus inactivation treatment. In a preferred embodiment, the cation exchange treatment is performed after solvent detergent virus inactivation. By this step, in addition to some undesired protein material (such as IgA, IgM and albumin) found in human serum and PEG, IgG from the residual reagents used for solvent-detergent treatment through cation exchange treatment Are separated.

If not otherwise achieved during the whole process, it should be subjected to a solvent-surfactant treatment in order to remove all particles which may also contain denatured proteins. Therefore, it is preferred to filter the solution with a 1 micron or finer membrane prior to solvent-surfactant addition. This will also reduce the virus-like material that may have come into the large particles and have avoided exposure to solvent-surfactants.

The filtrate is diafiltered and / or concentrated to a concentration of up to about 12% protein, preferably 5-10% protein, and for solvent or solvent-detergent treatment. Attached.

Today, the preferred solvent used for the inactivation of capsular viruses is trialkyl phosphate. The trialkyl phosphate used in the present invention is not particularly limited, but is preferably tri (n-butyl) phosphate (hereinafter “TNBP”).
Is. Other usable trialkyl phosphates are tri (ter-butyl) phosphate, tri (n-hexyl) phosphate, tri (2-ethylhexyl).
Phosphate, and others. It is also possible to use a mixture of two or more different trialkyl phosphates.

The trialkyl phosphate is 0.01 to 10 (w / w)%, preferably 0.1 to 3 (w / w)
)% Used.

Trialkyl phosphate is a surfactant or surfactant (s)
urfactant) in the presence or absence. Preferably, the trialkyl phosphate is used in combination with a surfactant. Detergents serve to contact the virus in the immunoglobulin mixture with the trialkyl phosphate.

As an example of the surfactant, polysorbate 80 (trade name: Tween 80, etc.)
And fatty acids including polyoxyethylene derivatives such as Polysorbate 20 (trade name: Tween 20, etc.), partial esters of anhydrous sorbitol, nonionic oil bath rinse agents such as oxidized ethylated alkylphenols (trade name: Triton X100, etc.) (Oil bath rinsing agent). Also included are surfactants and surfactants such as zwitterionic surfactants and others.

When using a surfactant, no critical amount is added, for example a concentration of about 0.001% to 10%
, Preferably between about 0.01% and 3%.

In the present invention, the trialkyl phosphate treatment of the mixture containing immunoglobulins is performed at about 20 to 35 ° C., preferably 25 to 30 ° C. for 1 hour or more, preferably about 5 to 8 hours.

During the trialkyl phosphate treatment, the aqueous medium of the protein solution contains about 5-10% immunoglobulin.

If not prior to solvent-detergent treatment, an optional anion exchange treatment can be performed on solvent-detergent treated immunoglobulins. Preferably, at least a cation exchange treatment is applied to the solvent-surfactant treated product. The ion exchange treatment is carried out on the aqueous immunoglobulin solution obtained in the solvent (or solvent surfactant) step, and usually has a pH of about 4.5 to 6.5 and the maximum amount of IgG is adsorbed at low ionic strength. The protein concentration is usually about 1-15 w / v%, more preferably between about 3 and 10 w / v%. The ion exchanger is equilibrated with the aqueous solvent used.

The continuous system is carried out by passing the immunoglobulin through the anion exchange column at a rate of 10 to 100 ml per ml of ion exchanger and collecting the non-adsorbed fraction.

The anion exchanger is used, for example, by binding an anion exchange group member to an insoluble carrier. The anion exchange group is diethylaminoethyl (DEAE),
There is a quaternary aminoethyl (QAE) group and others, and agarose, cellulose, dextran, polyacrylamide and others as insoluble carriers. One gram of DEAE Sephadex A-50 resin is swollen with 0.4% sodium chloride solution to a wet weight of 20 to 30 grams.

Cation exchangers that can be used are carboxymethyl sephadex (CM-Sephadex), CM-cellulose, SP-Sephadex, CM-Sepharose and SP-Sepharose. Using a pretreated cation exchanger (eg 1 gram of CM-Sephadex C-50 resin swollen with 0.4% sodium chloride solution to a wet weight of 25 to 35 grams) as a column bed, It is continuously passed through an immunoglobulin solution from a solvent (or solvent surfactant) at about 0-5 ° C.

The conditions described above are used for cation exchangers, IgG is adsorbed, then the protein-adsorbed cation exchange resin can be washed and the IgG can be eluted with, for example, 1.4N sodium chloride solution. .

When the ion exchange treatment is not adopted, the solvent (solvent surfactant) -treated solution is diafiltered and concentrated by tangential flow filtration to obtain the solvent surfactant. The agent and PEG are removed. Solvent surfactants and PEG are desired to be at very low levels in the final product and are therefore preferably anion exchanger followed by tangential flow filtration.

By the steps that follow the above steps, the IgG is clarified, diafiltered and concentrated to the required degree. If desired, a stabilizer such as D-sorbitol is added and the final recovery is 50-100 mg / ml IgG and 50 mg / ml D-sorbitol at pH about 5.4. . This solution may be passed through a membrane with a pore size of 35 nanometers or less for further virus removal. This stable and optionally nanofiltered solution is filtered through a sterile, bacteria-capable filtration membrane and filled into vials.

The following examples illustrate, but do not limit the invention.

[0053]

【Example】

  Method for producing intravenous immunoglobulin and resulting product

1800 grams of Fraction (Fr) II + III paste was suspended in 15 kg of cold water. 0
After mixing for 1 hour between 5 and 5 ° C, adjust the pH of the suspension to about 5.0 with diluted acetic acid.
Adjusted to. After the suspension was mixed for 3 hours, about 900 grams of filter aid (acid washed celite) was added and mixed for 45 minutes. Insoluble material was removed by filtration along with Celite. The filtrate was clarified and concentrated by ultrafiltration with a 100,000 molecular weight cutoff (MWCO) membrane to a protein concentration of approximately 6%.

D-sorbitol is added to the concentrated protein solution to a final concentration of 33 w / w% and mixed at pH 5.0 until all the sorbitol is dissolved. The solution was then heated at 60 ° C. for 10 hours. The heated solution was cooled to below 10 ° C and diluted with the same weight of cold water. The pH of the diluted solution was adjusted to 5.7 with diluted sodium hydroxide and 50% polyethylene glycol (PEG) 335.
0 was added to give a final concentration of 6 w / w%. Mix for about 2 hours at pH 5.7, and remove the precipitate formed by filtration, and wash with acid washed Celite filter aid 3 times.
% W / w was added. About 3.5 kg of 6% PEG filtrate was provided for another experiment. After adjusting the pH of the remaining 6% PEG filtrate to 4.9, bentonite was added in an amount of 1 g per kg of the filtrate to raise the pH to about 5.1. After mixing for 2 hours, the suspension was filtered to remove bentonite. Concentrate the filtrate and 100,000 MW
A diafilter was performed by ultrafiltration using a CO membrane.

The pH of the solution was adjusted to about 6.5 with diluted sodium hydroxide, and 0.4 kg of DEAE Sephadex A-50 resin previously expanded was added to the solution.
After mixing the protein solution with the resin, the resin was removed by filtration. Tri-n-
Solvent surfactant (SD) solutions containing butyl phosphate (TNBP) and polysorbate 80 were added to the filtrate at final concentrations of 0.3 w / w% and 1.0 w / w%, respectively. After the SD-containing solution was heated at 27 ° C. for 6 hours, the solution was cooled to 0 to 5 ° C., and the conductivity was adjusted to about 7 mS / cm and the pH to 5.8 with a sodium chloride solution.
About 2.7 kg of pretreated CM-Sephadex C-50 resin was added to the solution,
Mixed and filtered so that the resin remained. The resin containing the adsorbed IgG,
It was washed with 0.3% sodium chloride solution and the adsorbed IgG was eluted with 1.4 M sodium chloride solution. The eluate was clarified, concentrated and diafiltrated with cold water. D-sorbitol was added, and the final solution recovery was a composition containing about 100 mg / ml IgG and 50 mg / ml D-sorbitol at a pH of about 5.4. After the final adjustment, the solution is sterile filtered,
Fill glass vials. The test results of the resulting products are shown in the following table.

[0057]

[Table 1]

─────────────────────────────────────────────────── ─── Continued front page    (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, I T, LU, MC, NL, PT, SE), OA (BF, BJ , CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, K E, LS, MW, MZ, SD, SL, SZ, TZ, UG , ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, C H, CN, CR, CU, CZ, DE, DK, DM, DZ , EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, K G, KP, KR, KZ, LC, LK, LR, LS, LT , LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, RU, S D, SE, SG, SI, SK, SL, TJ, TM, TR , TT, TZ, UA, UG, UZ, VN, YU, ZA, ZW (72) Inventor Andranik Bagdasarian             United States California 91773               San Dimas Curry Estrella 1227 (72) Inventor Gorgonio Canaveral             United States California 91789               Walnut Silver Valley Tray             Le 608 F term (reference) 4C085 AA33 CC22 DD34 DD39 DD41                       GG02

Claims (21)

[Claims] 1. A continuous process for the preparation of an injectable gamma globulin solution comprising: (a) an impure gamma globulin solution under conditions of sufficient time and temperature to inactivate the heat-sensitive virus. (B) fractionating the heat-treated gamma globulin solution with polyethylene glycol without causing precipitation of the desired gamma globulin without recovering the product;
And (c) without recovering the product, the purified gamma globulin solution is treated with an organic solvent for inactivation of the coat virus. 2. The process according to claim 1, wherein the impure gamma globulin solution contains Cone fraction I + II + III, Cone fraction II + III paste, Cone fraction II + IIIw, or Cone fraction II. . 3. The process according to claim 1, wherein the impure γ-globulin solution contains a protein derived from the Cohn fraction II + III paste. 4. The impure γ-globulin solution is subjected to at least one step of purification treatment before the heat treatment means (a), and a heat treatment method without partially recovering the partially purified solution. The manufacturing process according to claim 1, which is attached to (a). 5. The process according to claim 1, wherein said heat treatment means (a) is carried out at about 50 to 70 ° C. for about 10 to 20 hours. 6. A process according to claim 5, wherein said heat treatment means (a) is carried out at about 60 ° C. for about 10 to 11 hours. 7. The process according to claim 1, wherein the one-step treatment step performed in the polyethylene glycol fraction comprises at least one means of removing impurities as a precipitate fraction and leaving the desired gamma globulin in solution. Process. 8. The process according to claim 6, wherein the impure gamma globulin solution contains a protein derived from the Cohn fraction II + III paste. 9. The process according to claim 1, wherein the organic solvent used in the means (c) is an alkyl phosphate. 10. The production process according to claim 7, wherein the organic solvent used in the means (c) is an alkyl phosphate. 11. The process according to claim 10, wherein the alkyl phosphate is tri-n-butyl phosphate. 12. The manufacturing process according to claim 1, wherein the organic solvent contains a surfactant. 13. The manufacturing process according to claim 11, wherein the organic solvent contains a surfactant. 14. The production process according to claim 1, wherein after the means (c), the γ-globulin solution is subjected to cation exchange treatment without being recovered. 15. The process according to claim 7, wherein the gamma globulin solution after the polyethylene glycol fractionation is subjected to bentonite refining means. 16. The process according to claim 15, wherein the bentonite treatment solution is treated with an anion exchange resin. 17. The process according to claim 1, wherein the gamma globulin solution after the step (b) is treated with an anion exchange resin. 18. The process according to claim 14, wherein the gamma globulin solution is concentrated by tangential flow filtration after the cation exchange treatment. 19. After the means (c), tangential flow filtration without recovering the gamma globulin solution.
The process according to claim 1, wherein the concentration is carried out by on). 20. An injectable gamma globulin solution produced by the manufacturing process according to claim 1. 21. An injectable gamma globulin solution produced by the manufacturing process according to claim 18.