CS239838B1 - Processing of humen imunoglobuline - Google Patents

Abstract

A method of preparing human immunoglobulins for intravenous administration anti-complementary activity and hypotensive the effect of immunoglobulins is reduced by incubation at -f-37 ° C and acidic pH presence of pepsin in the maltose environment. Maltose has anti-aggregation and stabilization effect on 1'human protein, thereby preventing undesirable structural changes in immunoglobulins. The presence of maltose also has a favorable effect affects the dissolution rate of the bot product. The nature of the human immunoglobulin production method for intravenous administration rests in that the ethanol-derived immunoglobulins fractionation in an environment of 0.1 to 5% maltose at pH 3.8 to 4.2 and temperature -j-37 incubated in the presence of pepsin for 18 to 25 hours. low molecular weight the substances are removed by dialysis versus 0.1 to 5% maltose solution. Post-dialysis dialysis appropriate p-parameters are sterilized by filtration, fill and lyophilize.

Description

The present invention is directed to methods for preparing human immunoglobulins for use in an intravenous application, which achieve a reduction in the anti-complementary activity and the hypotensive effect of immunoglobulins.

Nowadays, immunoglobulins used for li-intravenous application are prepared in the Czech Republic in such a way that immunoglobulins isolated by ethanol fractionation remove substances with anticomplementary activity and hypoteinsive effect, either by dialysis using dialysis intestines (Štěpánek)

I .: AO 202 180 and AO 208 86Š), or using ultrafiltration equipment with defined pore sizes.

The disadvantage of this product is long solubility time and possibility of reagregation of immunoglobulin molecules during the technological process, resp. during storage.

Abroad, to prepare immunoglobulins for intravenous administration, technological procedures for the cleavage of immunoglobulins by pepsin are used (Koch F .: Dtsch. Med. Wschr. 88: 282, 1963, Schultz, E., Schwick. G .: Dtsch. Med. Wschr. 87: 1643, 1922, optionally by repeating the immunoglobulin, by incubating at acidic pH in the presence of pepsin (Barandun S., Kistler P., Jeunet F., Istiker H., Vox Sang. 7, 1962, pp. 157-174).

A disadvantage of the compositions thus produced is altered biological properties, such as opsonization ability and other biological activities.

The essence of the novel process for the preparation of human immunoglobulins for intravenous administration is that a 5-8% solution of immunoglobulins containing 0.1 to 5% maltose at a pH of 3.8 to 4.2 and a temperature of 36 to 36% to 38 ° C is incubated in the presence of pepsin for 18-26 hours. After incubation, the pH of the rhombus is adjusted to 6.8 to 7.0 and then dialyzed against 0.1 to 7.0.

Product quality assessment Specified parameters appearance solubility antique. activity hypotensive effect (pressure drop)

A product prepared by an existing technol.

slight opalescence 30 ‘

0.71 j / mg bleach. —5.0% 0.0-0.1% Maltose Solution, Dlalysate after setting, the appropriate parameters of the filter are, via _; bacteriological filters, fill, freeze, and lyophilize.

The proposed process for the preparation of human immunoglobulins for intravenous administration removes undesirable properties of immunoglobulins, with care being taken in the technological process. that the structure of the immunoglobulins is impaired as little as possible.

Example 1

To 100 ml of a 5 to 8% solution of immunoglobulins containing 1.5% maltose, whose pH is adjusted to 3.8 to 4.2, add 50 milligrams of pepsin at an efficacy of 1: 110,000 and then incubate at a temperature of . -) - 37 ° C for 25 hours. After the incubation is complete, the pH of the solution is adjusted to 6.8 and dialyzed against a 0.1 to 5% maltose solution at - 1-4 ° C for 12 to 48 hours. After adjusting the protein content to 4.5-5.5%, the immunoglobulin solution is filtered through bacteriological filters, filled, frozen and lyophilized.

Example 1 and d 2

To 100 ml of a 5 to 8% solution of immunoglobulins containing 1.0% maltose whose pH is adjusted to 3.8 to 4.2, add 50 mg of pepsin with an efficacy of 1: 10,000 and then incubate at -j- 37 ° C for 18 hcidin. After incubation, the pH of the solution is adjusted to 6.8 and dialyzed against a 0.1 to 5% maltose solution at +4 ° C for 12 to 48 hours. After adjusting the% protein to 4.5-5.5, the immunoglobulin solution is filtered through bacteriological filters, filled, frozen and lyophilized.

Product prepared according to PV clear ·

2 '

0.08 j / mg bleach. .

Claims (2)

SUBJECT A process for the production of human immunoglobulins for intravenous administration, characterized in that a 5 to 8% immunoglobulin solution obtained by ethanol fractionation is in the presence of 0.1 to 5% maltose in the presence of OF THE INVENTION pepsin at acid pH incubates at 36-38 ° C for 18-25 hours. 2. The method according to claim 1, characterized in that the immunoglobulin solution by β-incubation is dialyzed against a 0.1 to 5% maltose solution for 12 to 48 hours.