DD160763A3 - PROCESS FOR THE PRODUCTION OF CONCENTRATE VACCINES - Google Patents

Abstract

The invention relates to a process for the preparation of concentrate vaccines from purified virus antigens and is intended to eliminate the existing defects with regard to unskillfulness, effectiveness and economic production. For the preparation of concentrate vaccines, the antigen is adsorbed in a closed system from a bacterially sterile solution for purification and concentration to a physiologically unshakable colloidal carrier, preferably calcium phosphate, and the adsorpt after isolation by sedimentation and decanting the supernatant liquid phase directly mixed with an effective adjuvant , Here, the antigen concentration can be adjusted to the desired value. The invention is predominantly suitable for the production of vaccines from immunogenic virus structural components or inactive virus.

Description

205 7 80

a) Title of the invention

Process for the preparation of concentrate vaccines

b) Field of application of the invention

The invention relates to a process for the preparation of concentrate vaccines, in particular of inactivated vaccines against foot-and-mouth disease (FMD). .

c) Characteristic of the known technical solutions

It is known that in inactivated virus vaccines the efficacy of the amount and purity of the inoculated em immunizing antigen, and the harmlessness of the type and amount of the accompanying substances depend · It has been demonstrated that especially antibiotics and tissue and cell components from the Propagating substrate of the virus are responsible for the occurrence of allergic reactions of the vaccinee. In order to be able to produce virus vaccines with sufficient purity and concentration of the immunizing antigen, methods are needed which allow an enrichment and purification of the immunizing antigen. Produce antigens under sterile conditions and are practicable on an industrial scale

It has already been proposed for the "concentration and purification of viral antigens on the industrial scale precipitation by Bolyäthylenglykol (MORROW, among others, 1974;. Bayra-MOGLU u _{e ε} β, 1974 ϊ LEI ₉ 1974) (barren polyethylene oxide ADAMOWICZ u" a " 1974). In these methods, the sediments of the viral antigens must be separated by centrifugation or filtration, whereby additional measures

205 780

There is also concern about the safe inactivation of viral aggregates resulting from the dispersion of the virus sediments. A technically attractive simple way of concentrating viral antigens will be used in the preparation of vaccine vaccines in which the virus is bound to the adsorbent simultaneously serving as Ad-juvans and the supernatant is removed by sedimentation of the adsorbent by decantation »When using aluminum hydroxide (MITBV'u.a.-, 1966) or'Bentonite (BAYiIAMOGLU u YALIM, 1974), the cleaning effect is unsatisfactory. Virus vaccines in which the antigen has been concentrated by adsorption on calcium phosphate (French Patent No. 122284) have unsatisfactory effectiveness,

d) Object of the invention

The aim of the invention is to eliminate the shortcomings in the production of concentrate vaccines with regard to harmlessness, effectiveness and economic production. "

e) Presentation of the essence of the invention

- The technical problem which is solved by the invention. -. '; , V

It is an object of the present invention to provide a technically simple process for producing concentrate vaccines from purified virus antigens in which the concentration and purification of the viral antigen and its further processing into finished vaccines are carried out under sterile conditions in a closed system without intermediate isolation steps "The vaccines prepared by this method should certainly not be infectious and cause no allergic reactions in the vaccinee and, if necessary after enrichment of the antigen, provide safe vaccination protection *

205 780

Features of the invention.

Bs it was found that one arrives at a technically easily feasible process for the preparation of concentrate vaccine ^ n from purified virus antigens, when the antigen from a bacterially sterile solution of a physiologically innocuous colloidal carrier, preferably calcium phosphate absorbed and the adsorpt after isolation by sedimentation and decanting the supernatant liquid phase directly with an effective adjuvant, whereby the antigen concentration can be adjusted to the desired value. "

Bacterially sterile solutions of the viral antigen which are free of cell debris and cell or viral aggregates capable of impairing safe inactivation of virulent viral antigens can optionally be prepared by filtration or homogenization with organic solvents that are immiscible with water. Adsorption the viral antigen on the colloidal carrier for purification and concentration of the immunizing antigen is advantageously carried out as adsorption precipitation at low temperatures, the colloidal carrier being formed in the virus solution itself by adding the solutions.

In this case, the reaction conditions (concentration of the adsorbent, pH and ion concentration of the medium) according to the characteristic properties of the antigen and the nature and concentration of the accompanying substances to modify in order to achieve a quantitative adsorption <of the antigen and an optimal cleaning effect. During the Adsorptionsfällung possibly auftre- tende undesirable changes in the pH of the virus solution can be advantageously avoided by the solution used for the precipitation of multivalent cations with an amount of an alkaline substance, preferably tris or sodium hydroxide is added, that during the Reaction resulting hydrogen ions are neutralized,

 When using virulent virus as an antigen can

ί U5 / 80

its inactivation by means of chemical or physical methods optionally before or after adsorption to the colloidal carrier or preferably after mixing with the adjuvant under optimal conditions (pH and temperature); be performed.

Mixing the virus antigen attached to the colloidal carrier with an adjuvant mixture, preferably aluminum hydroxide, saponin or DBAE-dextran, results in virus vaccines that are non-infectious and free of allergy-causing substances. By mixing the antigen adsorbed to the colloidal carrier, preferably calcium phosphate, with an additional adjuvant, a combination with increased adjuvant effect is produced. The efficacy of the vaccines prepared in the manner described is greater than that of vaccines in which untreated antigen has been processed with the same adjuvants. The method described herein for the production of concentrated vaccines from purified viral antigens can be distinguished from the provision of the sterile clarified or pre-cleaned virus solution to fill the inactivated vaccine in a closed system without additional technological operation beyond going over the sterile solutions or suspensions realize.

f) embodiments

1 «Preparation of a purified trivalent vaccine against foot-and-mouth disease.

1125 g of Ka ₂ HBX χ 2HgO are dissolved in 200 l of water and sterilized by autoclaving. 500 l of a mixture of MKS cell culture virus from 3 serological types, which have been partially purified by chloroform homogenization and subsequent centrifugation and germ-free, are added to the phosphate solution and cooled to a temperature 10 ⁰ C. By adding 60 l of a Tris-buffered CaCl ₂ solution (0.1 M CaCl ₂ , 0.025 M Tris), hydrated calcium phosphate is precipitated in the solution, which adsorbs the virus from the solution. By adding

205 780

Acid or alkali must be ensured that a pH of 7.3 is maintained during the adsorption phase. After sedimentation of the adsorbent at low temperatures, the clear supernatant with the non-adsorbed by-products is completely removed and the sediment is mixed with 900 l of a sterile 1% suspension of aluminum hydroxide in a 0.07 m buffered UaCl solution which is 1.500 The suspension is treated with enough UaOH so that the finished vaccine has a pH of at least 8.2 after setting a formaldehyde content of 190 mgA and filling to a total volume of 1000 l Virus is 24 hours at 25 ° C.

2, production of a monovalent concentrate vaccine against foot-and-mouth disease with increased antigen content,

500 l of a serological type MKS cell culture virus are purified and concentrated under the same conditions as described in FIG. 1, embodiment. The adsorbent is mixed with 150 g of saponin and 160 g of glycine and inactivated after filling to 100 1 at pH 8.2 and a formaldehyde content of 190 mgA for 24 hours at 25 ° C.

¹ . Y

ι Π H \\ l |: Γ / ν ίί * 'ι ^{": ·} ί

Claims (2)

invention claims 1. A process for the preparation of concentrate vaccines from bacterially sterile solutions of active or inactive virus antigens, which are free of cell debris and cell or virus aggregates, characterized in that the antigen for the purpose of purification and concentration of a physiologically harmless carrier, preferably colloidal calcium phosphate, is adsorbed and the separated by sedimentation Adsorpt is mixed directly with an adjuvant or an adjuvant combination, whereby by appropriate selection of the volume ratios a predetermined concentration of the virus antigen is set « 2, method according to point 1, characterized in that in the adsorption precipitation with calcium phosphate Kalziura , Chloride solution, which is added to generate the precipitate in the solution containing an equivalent amount of sodium phosphate added virus solution, with such an amount of an alkaline reactant, preferably tris or sodium hydroxide is mixed, that the resulting during the reaction of hydrogen ions are neutralized