Biotest-Serum-Institut Gmbh

Biotest Pharma Gmbh

Boitest Pharma Gmbh

Ronald Kotitschke

Wolfgang Stephan

Dieter Rudnick

Wolfgang Moller

Axel W Stemberger

Detlef Piechaczek

A method of preparing a sterile and stable plasma-protein solution containing fibrinogen and Factor XIII from human blood plasma which has been stabilized with citrate, comprising treating the plasma with .beta.-propiolactone and irradiating it with ultraviolet light, removing the Factors II, VII, IX and X by adsorption onto anion exchangers that adsorb proteins, precipitating the companion proteins out by adding ethanol until the solution has a final concentration of about 9% by volume at -3.degree. C., centrifuging the precipitate off, dissolving the precipitate in a citrate buffer at a pH of about 6.35 and a temperature of about 37.degree. C., adjusting the protein level of the solution to about 13.3 g/l with sodium citrate solution, adding ethanol, a glycine citrate buffer, and a solution of sodium citrate to precipitate out the companion proteins, adding ethanol to the remaining solution until the solution has a final concentration of about 9% by volume at -3.degree. C., thereby precipitating fribrinogen and Factor XIII, dissolving the precipitate in a citrate buffer at a pH of about 6.35 and a temperature of about 37.degree. C., and filtering the solution.

Method of preparing a sterile plasma-protein solution containing

Process for the preparation of an immunoglobulin solution suitable for intravenous administration from a human blood protein fraction containing immunoglobulins IgG, IgA and IgM in partially concentrated from, with the process steps: addition of acetate buffer to the protein fraction, where appropriate removal of insoluble constituents by filtration, treatment with calcium phosphate and octanoic acid, centrifugation, removal of the supernatant and treatment thereof with an adsorbent, removal of the adsorbent and sterilization by filtration.

Process for the preparation of a pharmaceutical which contains IgG, IgA

To maintain an almost unchanged plasma-protein profile in a patient subsequent to plasma exchange, the plasma-exchange medium contains the most essential human serum proteins, except for the coagulation factors, at a concentration of 75 g/l.

Replacement of human plasma using sterile solution of human plasma

A coagulant plasma-protein solution obtained by adsorbing stabilized human plasma at least once with 20-40 mg of colloidal silicic acid per gram of plasma protein and separating the adsorbent from the adsorbate, a method of manufacturing the solution, and a pharmaceutical preparation containing the solution and intended for the intravenous treatment of complex disorders of the hemostatic system. The solution is low in fibrinogen and factors XI and XII but otherwise close in composition to the starting material.

Coagulant plasma-protein solution

A process for the cold sterilization of a preparation containing blood coagulation factor VIII, comprising

Method for the cold sterilization of preparations containing blood

The present invention relates to a method for making fibrinogen, a prothrombin complex containing the coagulation factors II, VII, IX and X that can contain antithrombin III, antithrombin III and a solution of stable serum proteins from a blood plasma stabilized with citrate which is characterized in that from the plasma, by adsorption on colloidal silica of a specific surface of 50 to 400 m.sup.2 /g and a concentration of 50 to 400 mg per g plasma protein, fibrinogen is isolated; and in that thereupon (a) citrate and calcium ions are removed by ultrafiltration or dialysis and then from the protein solution, over anion exchangers or tricalcium phosphate that adsorb proteins, the coagulation factors II, VII, IX and X and antithrombin III are adsorbed, or (b) the coagulation factors II, VII, IX and X are adsorbed prior to the ultrafiltration or dialysis, antithrombin III then not being simultaneously adsorbed, and antithrombin III is adsorbed on the said adsorbents after the removal of the citrate and calcium ions by ultrafiltration or dialysis; and in that thereupon from the remaining plasma fluid further unstable proteins are removed by another adsorption on colloidal silica and a solution of stable serum proteins is obtained.

Fractionating citrate-stabilized plasma

A citrate-stabilized blood plasma is frozen and thawed, the undissolved cryoprecipitate is separated and is rich in factor VIII protein. The residual liquid is treated with cation and anion exchangers, either sequentially or in admixture, to separate calcium and citrate ions therefrom. The residual plasma is then subjected to adsorption to remove a fraction which is eluted and precipitated, enriched in factors II, VII, IX and X. The liquid left from adsorption is subjected to further adsorption with silicic acid, leaving a storage-stable serum. The silicic acid is eluted to dissolve adsorbed fibrinogen which then is re-precipitated.

Ion exchanger treatment of citrate-stabilized plasma

An antiserum for the quantitative determination of X and Y degradation products of fibrin and fibrinogen is produced by administering the purified degradation products to an experimental animal thereby to immunize said animal, drawing serum from said animal, contacting said serum with a plasma and separating the resultant precipitate, thereby to leave a supernatant liquid which constitutes said antiserum. The plasma may be a heparin, citrate, oxalate, ethylenediaminetetracetate or glutaraldehyde-treated plasma and as many as 12 or 15 precipitations may be made with the animal antiserum. The antiserum may be fractionated at any stage by adsorption on Sephadex modified dextran.

Preparation of antiserum for quantitative determination of X and Y

A process for the fractionation of blood comprising passing said blood through a cation exchanger, separating the solids from the plasma, freezing the plasma, thawing the frozen plasma, and separating a first product comprising undissolved cryoprecipitate enriched in factor-VIII protein from plasma fluid. The cryoprecipitate can be concentrated by warm water dissolution and polyethylene glycol precipitation. The plasma fluid, optionally after treatment with .beta.-propiolactone and uv irridiation, is treated with tricalcium phosphate to adsorb factors II, VII, IX and X as a second product. These factors can be eluted with citrate solution. The residual plasma from the initial tricalcium phosphate adsorption is treated with colloidal silica to adsorb impurities and leave a third product which is a storage-stable serum protein solution.

Growing community of inventors

Dreieich, Germany

Ronald Kotitschke