Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2/00—Disinfection or sterilisation of materials or objects, in general; Accessories therefor
  • A61L2/16—Disinfection or sterilisation of materials or objects, in general; Accessories therefor using chemical substances
  • A61L2/18—Liquid substances
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/14—Blood; Artificial blood
  • A61K35/16—Blood plasma; Blood serum
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2103/00—Materials or objects being the target of disinfection or sterilisation
  • A61L2103/05—Living organisms or biological materials

Definitions

• the present invention relates to a method for producing non-infectious blood plasma.
• Blood plasma is used in medicine for an abundance of indications. Its clinical importance is correspondingly great.
• the blood plasma is obtained by separating the blood cells from whole blood that has been taken from donors. In order to avoid infections of the patient with pathogenic components, such as hepatitis viruses and HIV of the donor plasma, these must be free of intact viruses. For this reason, it was previously only possible to use plasmas from individual donors which, after a longer observation period, could be considered healthy in the sense of being virus-free. Mixing (pooling) plasmas from several donors was excluded.
• a disadvantage of this procedure is the enormous outlay which must be expended by the individual preparations. A continuous way of working is not possible with the currently existing working techniques. As a result, a further disadvantage appears. It is only possible with disproportionately high, hardly justifiable expenditure to produce standardized plasma preparations. Due to the individual preparation, for example, the protein content in the donor fraction fluctuates. Therefore, when using the plasma produced by individual preparation, the current protein content of the batch in question must be taken into account. A major problem is the removal of the agent used to inactivate the infectious components. According to a procedure developed by the New York Blood Center, detergents are used for virus inactivation. A major problem is to remove these detergents from the plasma after virus inactivation. Horowitz suggests removing the detergent-treated preparations from the detergent using hydrophobic chromatography using reverse phase materials.
• the technical problem on which the invention is based is therefore the provision of a continuous process for producing a non-infectious blood plasma.
• the problems resulting therefrom relate, among other things, to a safe and reliable procedure for removing the virus-inactivating detergents.
• the technical problems on which the invention is based are solved by a method for producing non-infectious blood plasma, the plasma being treated with non-ionic surfactants, the lipid phase being separated off and the non-ionic surfactant being removed by means of solid-phase extraction on hydrophobic materials.
• the blood plasma can originate from freshly prepared plasma preparations as well as from frozen fractions. Frozen plasma is thawed in a water bath. As soon as a certain temperature, advantageously being preferably 25 ⁇ C, is reached, the plasma is optionally filtered and the pH of the solution checked. The thawing process is preferably carried out in a time of about 1 to 2 hours.
• the pH of the solution is preferably 7.0 to 7.4.
• the blood plasma treated in this way is then treated with a mixture of nonionic surfactants and distilled water at 20 to 40 ° C., preferably 30 ° C.
• the nonionic surfactants used are in particular tri-N-butyl phosphate (TNBP) and / or Triton X-100. It is advantageous to treat the mixture for a while at elevated temperature with gentle stirring.
• the lipid fraction is then removed by centrifugation. Before the centrifugation step, it is advantageous to treat the lipid fraction with biologically compatible lipids In a preferred embodiment of the method according to the invention, about 10% vegetable oil is added. After centrifugation, the lipid and plasma layers are separated. This can preferably be done by means of a siphon or a continuous one working centrifuge with automatic phase separation happen.
• the plasma layer is collected while the lipid layer is discarded. This is followed, if appropriate, by a filtration step, whereupon the remaining non-ionic detergent which is still present is removed from the plasma by means of solid phase extraction.
• the hydrophobic material for solid phase extraction is preferably the material known from reversed phase chromatography. These preferably include carrier materials coated with C-18, as are also used in high-pressure liquid chromatography. In a preferred embodiment the solid phase extraction can be carried out in the form of a chromatography.
• the ratio between bed volume and the plasma volume to be processed is preferably 1: 6 to 1:20, particularly preferably 1:10.
• the pH may be checked.
• the pH of the solution is readjusted to pH 7.0 to 7.4. Then one closes.
• Sterile filtration whereupon, if desired, the blood plasma is freeze-dried in the presence of customary lyophilization aids, such as lysine or glycine.
• the method according to the invention makes it possible to depart from the customary individual preparation of the plasma.
• the process is suitable for being carried out on an industrial scale.
• a continuous procedure is made possible.
• a further advantage of the process according to the invention is that, because of the continuous process design, the conditions can be selected so that the end product has the same or at least very similar protein concentrations. This makes the clinical handling of the blood plasma preparations, especially in the case of multiple doses, less problematic, since the changing amount of protein resulting from the individual preparations is eliminated.
• 20 1 blood plasma (80 x 250 ml) are thawed in a water bath at 25 ° C for 1.5 hours. If desired, the fraction is thawed over gauze filters or filter cartridges (eg company Pal, 100 ⁇ m) filtered. The plasma is then filled into a 50 l stainless steel kettle at 30 ⁇ c. A propeller blade stirrer is used for stirring operations. The pH of the thawed plasma is checked and adjusted to pH 7.0 to 7.4 with 0.5 N hydrochloric acid.
• a mixture of TNBP, Triton X-100 and distilled water is produced.
• 200 ml TNBP, 200 ml Triton was used in a particularly preferred manner X-100 and 800 ml of distilled water were used.
• the mixture is also heated to approx. 30 ° C.
• the mixture of non-ionic surfactants is added to the plasma.
• the mixture is carefully stirred at 30 ° C. After 4 hours, the plasma is centrifuged. Centrifugation is preferably carried out at 4,000 revolutions per minute at 12 ° C. for one hour.
• This amount is treated with "C-18 preparative" adsorbent (Waters, Inc.). With the amount of 19 l obtained, 1.2 kg are preferably slurried in 2.4 l of water.
• the chromatography is carried out on a column with a diameter of 15 cm. The flow rate is 300 ml / min. The eluate is monitored using a UV monitor. The emerging plasma fraction is collected and, if necessary, adjusted to a pH of 7.0 to 7.4 with 0.5 N sodium hydroxide solution.
• the plasma is sterile filtered through membrane filters (0.22 ⁇ m) and freeze-dried after it has been filled into 250 ml bottles.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Cell Biology (AREA)
• Chemical & Material Sciences (AREA)
• Hematology (AREA)
• Epidemiology (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Biotechnology (AREA)
• Medicinal Chemistry (AREA)
• Engineering & Computer Science (AREA)
• Biomedical Technology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Developmental Biology & Embryology (AREA)
• Immunology (AREA)
• Virology (AREA)
• Zoology (AREA)
• General Chemical & Material Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Medicines Containing Material From Animals Or Micro-Organisms (AREA)
• Peptides Or Proteins (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Treatment Of Liquids With Adsorbents In General (AREA)
• Apparatus For Disinfection Or Sterilisation (AREA)
• Compounds Of Unknown Constitution (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Priority Applications (3)

Applications Claiming Priority (2)

Publications (1)

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ID=6402597

Family Applications (1)

Country Status (8)

Cited By (6)

Families Citing this family (6)

Citations (5)

Family Cites Families (2)

• 1990
  • 1990-03-20 DE DE4008852A patent/DE4008852A1/de active Granted
  • 1990-07-06 DE DE4021542A patent/DE4021542C2/de not_active Expired - Fee Related
• 1991
  • 1991-03-16 ES ES91906410T patent/ES2059129T3/es not_active Expired - Lifetime
  • 1991-03-16 WO PCT/EP1991/000503 patent/WO1991014439A1/de not_active Ceased
  • 1991-03-16 AT AT91906410T patent/ATE110272T1/de not_active IP Right Cessation
  • 1991-03-16 EP EP91906410A patent/EP0472708B1/de not_active Revoked
  • 1991-03-16 JP JP3506288A patent/JP2708632B2/ja not_active Expired - Lifetime
  • 1991-03-16 DE DE59102623T patent/DE59102623D1/de not_active Revoked
  • 1991-03-16 DK DK91906410.5T patent/DK0472708T3/da active
  • 1991-11-11 FI FI915301A patent/FI101128B/fi active

Patent Citations (5)

Non-Patent Citations (1)

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