JPH0665651B2 - Method for producing intravenous immunoglobulin preparation - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial field of use) Immunoglobulins are widely effective against infectious diseases such as agammaglobulinemia, rubella, vaccinia, and diphtheria, and are manufactured and sold as intravenous injection preparations. This immunoglobulin is mainly produced from corn fraction II, which is a gamma globulin fraction obtained from human plasma by the corn ethanol fractionation method. The present invention uses this corn fraction
It improves the method of producing immunoglobulin drugs from II.

(Prior Art) Immunoglobulin spontaneously aggregates during storage of human plasma, or aggregates by the action of drugs such as alcohol, acid and alkali in the separation, purification and commercialization process, and has an anti-complementary action in the aggregate. Demonstrate blood pressure drop, body temperature rise,
Causes side effects such as cardiovascular disorders. Therefore, conventionally, this aggregate was removed from the corn fraction II by polyethylene glycol treatment or the like before commercialization.

(Problems to be Solved by the Invention) However, this method has complicated steps, and
There is a problem in that the stability of the product obtained through this treatment is low.

(Means for Solving the Problems) The inventors of the present invention have earnestly studied to develop a method for easily obtaining an intravenous immunoglobulin preparation with low anti-complementary value and high stability. An immunoglobulin preparation for axial injection with low stability and low anti-complement number by efficiently removing aggregates and the fractions that tend to form aggregates when the solution containing II is passed through a hollow fiber made of polymethylmethacrylate The present invention has been completed based on the finding that it can be easily obtained.

That is, the present invention relates to a method for producing an intravenous immunoglobulin preparation from corn fraction II, wherein a hollow fiber made of polymethylmethacrylate is prepared from corn fraction II to which 0.5 to 2% of albumin or corn fraction V is added. The present invention relates to a method for producing an immunoglobulin preparation for intravenous injection, which comprises using an immunoglobulin solution.

Cohn fraction II is a gamma globulin fraction obtained by the Cohn ethanol fractionation method. Various modified methods have been developed for the corn ethanol fractionation method, but the kind does not matter. This corn fraction II may be a freeze-dried product,
Gamma globulin aggregates and dimers are produced during freeze-drying, so rather it is preferable to use the paste as a raw material.

An immunoglobulin solution that is filtered with a hollow fiber made of polymethylmethacrylate is a solution in the process from the corn fraction II to the production of an immunoglobulin preparation for intravenous injection. It is a solution dissolved in saline or an appropriate buffer solution. The buffer solution has a pH of about 4-9 and 5-200 mM.
A phosphoric acid type, citric acid type, acetic acid type or the like may be used. An immunoglobulin concentration of about 1 to 10% is suitable. By adding about 0.5 to 2% of albumin, which may be corn fraction V, to this solution, it is possible to enhance the stability of immunoglobulin during filtration and subsequent freeze-drying with a hollow fiber made of polymethylmethacrylate. it can.

As the membrane capable of separating plasma from blood, those for plasma separation therapy can be used. There are various materials for this film, such as polymethylmethacrylate, cellulose acetate, polyvinyl alcohol, and the like, and any type can be used.

The operating conditions may be in accordance with the normal conditions of the apparatus equipped with the above polymethylmethacrylate hollow fiber. Since immunoglobulin is permeated and collected through the hollow fiber made of polymethylmethacrylate, it is preferable to put the above-mentioned liquid for immunoglobulin solution in the circulation system on the permeation side. It is also effective to appropriately supplement the stock solution with the solution used for the immunoglobulin solution in the latter half of the incubation. The filtration with the hollow fiber made of polymethylmethacrylate may be performed in a batch system or a continuous system. In the case of the continuous system, it is preferable to appropriately draw not only the permeate side but also the stock solution side.

The permeate may be used as a liquid formulation as it is or by adding a stabilizer such as amino acid, or may be freeze-dried to obtain a product. Aseptic filtration, which has been generally performed conventionally, is usually unnecessary in the method of the present invention.

(Action) Monomers of immunoglobulin permeate the membrane, while aggregates do not permeate and remain. The dimer may permeate or may remain depending on the type of membrane.

Example 1 100 g of lyophilized powder of corn fraction II was dissolved in 50 mM sodium phosphate buffer pH 7.0, and 10 g of lyophilized powder of corn fraction V was dissolved to make a total amount of 1, Was used as the stock solution.

The membrane is made of polymethylmethacrylate hollow fiber (Plasmax
AS-08, a product of Toray Industries, Inc. was used, and a test was conducted using the apparatus outlined in FIG. This apparatus is composed of a filtration apparatus 1 equipped with a hollow fiber membrane, a stock solution storage tank 2, a stock solution side line 3 for supplying and circulating the stock solution to the excess apparatus 1, a peristaltic pump 4 for feeding the solution, and a permeate for permeation solution circulation. It comprises a liquid side circulation line 5 and a peris pump 6 for feeding the same, a permeated liquid drawing line 7 and its receiving tank 8.

The circulation rate of the stock solution is 100 ml / min, and the withdrawal rate of the permeate is
It was set to 10 ml / min.

The protein concentration of the permeate increases with increasing liquid volume,
The maximum value was reached when 600-700 ml was permeated. About 900 ml can be collected if the stock solution is left as it is, but the buffer solution used for preparing the stock solution should be added to keep the volume of the stock solution constant from the time when the protein concentration reaches the maximum value of 600 to 700 ml. While continuing to pass, the operation was stopped when the amount of permeated liquid reached 1800 ml.

The recovered 1800 ml of permeate was freeze-dried to obtain a freeze-dried product with a gamma globulin yield of 41% based on the stock solution. When the ratio of the gamma globulin monomer, dimer and aggregate of the stock solution, permeate and lyophilized product was measured, the following results were obtained.

The anti-complement value before freeze-drying was 10 units, whereas the anti-complement value of the freeze-dried product was 13 units.

The anti-complement value was determined by Kabat and Meyer's method (Mayer, MM: Experimental Immunochemistry (Ed. By K.
abat, EA & Mayer, MM), 2nd.Ed., pp133〜240 (196
1)).

Example 2 50 g of lyophilized powder of corn fraction II was dissolved in 50 mM sodium phosphate buffer pH 7.0, and 10 g of lyophilized powder of corn fraction V was dissolved to make the total amount 1.

When this was used as a stock solution and treated in the same manner as in Example 1, a lyophilized product was obtained with a gamma globulin yield of 45% based on the stock solution. When the ratio of the gamma globulin monomer, dimer and aggregate of the stock solution, permeate and lyophilized product was measured, the following results were obtained.

The anti-complement value before freeze-drying was 9 units, whereas the anti-complement value of the freeze-dried product was 12 units.

(Effects of the Invention) By using the method of the present invention, a product can be easily obtained even from a lot containing many aggregates. The residual liquid obtained by recovering gammaglobulin by the method of the present invention can be used again for filtration or enzyme treatment with a hollow fiber made of polymethylmethacrylate, and as a result, the recovery of gammaglobulin is very high. Become. Moreover, the product obtained by this method has low anti-complement value and high stability. The method of the present invention, after filtration with a hollow fiber made of polymethylmethacrylate,
The sterilized preparation that can be injected intravenously can be obtained immediately by lyophilization, so the operation is simple.

[Brief description of drawings]

FIG. 1 is an explanatory view showing an outline of an example of a filter device used in the method of the present invention.

Continuation of front page (56) Reference JP-A-56-59716 (JP, A) JP-A-56-81521 (JP, A) JP-A-56-59716 (JP, A) JP-A-49-81519 (JP , A) JP 59-116223 (JP, A) JP 49-101516 (JP, A) JP 53-81613 (JP, A) JP 59-196794 (JP, A) JP 61-12627 (JP, A) JP-A-56-81521 (JP, A)

Claims (1)

[Claims] 1. A method for producing an immunoglobulin preparation for intravenous injection from corn fraction II, which comprises adding a 0.5 to 2% albumin or corn fraction V to corn fraction II to obtain a polymethylmethacrylate hollow fiber. A method for producing an immunoglobulin preparation for intravenous injection, which comprises passing immunoglobulin using