WO2006122476A1 - A method of inactivating virus, treatment system and device used thereof - Google Patents

A method of inactivating virus, treatment system and device used thereof Download PDF

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Publication number
WO2006122476A1
WO2006122476A1 PCT/CN2006/000877 CN2006000877W WO2006122476A1 WO 2006122476 A1 WO2006122476 A1 WO 2006122476A1 CN 2006000877 W CN2006000877 W CN 2006000877W WO 2006122476 A1 WO2006122476 A1 WO 2006122476A1
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WO
WIPO (PCT)
Prior art keywords
medium
virus
virus inactivating
adsorption
organic solvent
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PCT/CN2006/000877
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French (fr)
Chinese (zh)
Inventor
Fanglin Zou
Chunsheng Chen
Jianxia Wang
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Chengdu Kuachang Medical Industrial Limited
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Filing date
Publication date
Priority claimed from PCT/CN2005/001397 external-priority patent/WO2006116899A1/en
Priority claimed from CN 200610008479 external-priority patent/CN1820786B/en
Application filed by Chengdu Kuachang Medical Industrial Limited filed Critical Chengdu Kuachang Medical Industrial Limited
Priority to CN200680013880XA priority Critical patent/CN101163510B/en
Publication of WO2006122476A1 publication Critical patent/WO2006122476A1/en

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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/22Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS 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
    • A61L2/00Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0082Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using chemical substances
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00Drugs for disorders of the blood or the extracellular fluid

Definitions

  • the present invention relates generally to methods of treating biological fluids, and more particularly to virus inactivation, treatment systems, and devices.
  • the virus inactivation especially the virus-killing activity containing the solid-liquid phase reaction, is taken as an example to illustrate the problems to be solved in the prior art.
  • the existing virus inactivation technique containing solid-liquid phase reaction is mainly based on organic solvent virus inactivation (Solvant-Detergent treatment, referred to as SD treatment in the present invention).
  • the organic solvent virus inactivating medium contains an organic solvent S (which may additionally contain detergent D) and a virus inactivating agent adsorbent such as activated carbon (refer to Chinese Patent 01123658.2).
  • virus inactivation techniques containing solid-liquid phase reactions including immobilization of alkylphosphorus compounds (using inorganic adsorbents as adsorbents, reference to German patent DE40 (H099A1), immobilized iodine treatment (with silicon-containing dry soil PVPP)
  • the filter plate is used as an adsorbent, refer to the product introduction of the German Shenk company, etc.
  • the problem of the prior art is that the activated carbon, the inorganic adsorbent or the silicon-containing dry soil PVPP filter plate has side effects (for example, strong protein adsorption capacity, Inactivation of coagulation factors, a significant increase in plasma APTT values, etc.).
  • the organic solvent virus inactivating agent adsorption medium is an inorganic adsorbent (for example, a perlite filter plate containing perlite:), an affinity adsorbent (for example). C-18 oleophilic resin), activated carbon.
  • the affinity adsorbent is expensive and not suitable for single use, and the activated carbon and inorganic adsorbent used have side effects (e.g., strong protein adsorption capacity, inactivation of coagulation factors, and substantial increase in plasma APTT value, and many more).
  • the photosensitizer virus inactivating agent adsorption medium is activated carbon (for example, PCT/US94/U227; US6159375; US08/347564), and Ion exchange resin (WO9103933), poly styrene-divinylbenzene, acrylic ester polymers (US6348309).
  • activated carbon, silica, and ion exchange resins have side effects (e.g., strong protein adsorption capacity, a large increase in plasma APTT value, etc.), while macroporous adsorption resins tend to fall off and contaminate biological fluids. Side effects can also be formed.
  • the expression "effective virus inactivating function" means
  • the expression "effective virus inactivating agent removal function” means a function of removing more than 90% of one or more virus inactivating agents.
  • the basic technical solution of the method of the present invention is to achieve the object of the present invention by using a solid phase medium having both effective functions and only minimal side effects.
  • the technical solution includes an embodiment: 1. In general, the addition of a oximation agent to a solid phase medium minimizes side effects in virus inactivation; 2. For a particular virus inactivating agent, a selection function/ Side effects can be achieved for the same purpose as the maximum solid phase medium.
  • the virus inactivation method of the present invention BJ comprising at least: ⁇ ) providing a biological fluid; ⁇ ) providing a treatment system; and C) contacting the biological fluid with the treatment system, wherein the treatment system comprises at least a solid a phase medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, the virus inactivating medium comprising a virus An inactivating agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: (a) a passivation adsorption medium, the passivation adsorption medium containing at least an adsorption medium and being fixed in adsorption a passivating agent on the medium; or/and (b) a physical adsorption medium, the physical adsorption medium comprising: (i) a photosensitizer physical ad
  • the treatment system of the present invention is the treatment system described in the above virus inactivation method of the present invention.
  • the processing system comprises at least a solid phase medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium,
  • the virus inactivating medium comprises a virus inactivating agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: (a) a passivation adsorption medium, the passivation adsorption medium being at least a passivating agent comprising an adsorbent medium and immobilized on the adsorbent medium; or/and (b) a physical adsorbent medium comprising: (i) a photosensitizer physics used as a photosensitizer virus inactivating agent adsorption medium Adsorbing fibers, the photosensitizer
  • the device of the present invention comprises at least the above described processing system.
  • Embodiments of the invention will illustrate that the objects of the invention can be attained by the method, processing system or apparatus of the invention. It is to be noted that the method, treatment system or device of the present invention is particularly suitable for the case where the solid phase medium is used at one time, in addition to the advantages of its small side effects.
  • biological liquid means a liquid containing or possibly containing a biological substance such as blood and blood components, plasma and plasma derivatives, biological products, biopharmaceuticals, and the like.
  • single human blood component means blood collected from one or more times from the same person or/and components separated therefrom, including single blood, single plasma, single serving. Platelets, single red blood cells, and more.
  • the term "effectively reducing pathogen harm” means a treatment process including effective virus inactivation or/and leukocyte removal;
  • the term “viral inactivation” means by adding a virus inactivating agent (for example)
  • SD agent photosensitizer virus inactivating agent, etc.
  • physical energy eg heat, light, etc.
  • a process for effectively reducing the infectious activity of at least a portion of the virus present in the preparation the term “white blood cell removal” "Includes a process for reducing leukocyte concentration by mechanisms such as adsorption and size filtration of leukocytes; the expression “effective” means, for virus inactivation, 99.9% or more of one or more model viruses are inactivated, For leukocyte removal, it means that more than 90% of white blood cells are removed.
  • virus inactivation treatment refers to the entire process of bringing a virus inactivating agent into contact with a biological fluid until the viral inactivating agent in the biological fluid is substantially removed.
  • Viruses include membrane viruses or/and non-lipid envelope viruses.
  • the virus inactivation treatment contains a virus inactivation process, and in many cases, a virus inactivating agent removal process. In the production process, the virus inactivation treatment and other treatments are performed sequentially or/and simultaneously, and thus the above virus inactivation treatment may also partially undergo or/and be accompanied by other treatments other than virus inactivation (eg, protein) Purification, etc.).
  • the term "viral inactivating agent” means any additive which has a virus inactivating function for all or part of viruses which may be present in a biological fluid, but may not be limited to a virus inactivating function, such as a photosensitizer virus inactivating agent/ Photosensitizer virus inactivating agent in light treatment, organic solvent virus inactivating agent in SD treatment or organic solvent virus inactivating agent/detergent, and the like.
  • the virus inactivating agent may have other functions besides the virus inactivating function.
  • psoralen is inactivated by viruses, but also inactivated by some other organisms (such as bacteria), in which case the virus inactivating agent is a bacterial inactivating agent, and the like.
  • viral inactivation agent "Biological” is a derivative of a virus inactivating agent formed in a virus inactivation treatment, such as a photosensitizer virus inactivating agent/a derivative of methylene blue which is a virus inactivating agent during photoactivation (for example, azure) A and B).
  • the organic solvent virus inactivation comprises virus inactivation by a combination of an organic solvent virus inactivating agent (S) or an organic solvent virus inactivating agent and a detergent combination (SD) for the virus inactivating agent.
  • the SD treatment includes an SD treatment method invented by the New York Blood Center (US Pat. No. 4,540,573), and a derivative method developed by others (for example, a virus-in addition method in which only S is added, and a virus inactivation treatment method in which S is shared with alcohol, etc.).
  • the term "organic solvent virus inactivating agent (abbreviated as S in the present invention)" means an organic solvent used under specific conditions of virus inactivation treatment.
  • reaction kinetic conditions are less favorable (eg, liquid phase)
  • concentration of the virus inactivating agent when the virus is inactivated is generally not more than 1% (w/v) ; the reaction conditions are relatively conservative (for example, the reaction temperature is usually less than 38 ° C and the pH is between 4 and 10); Therefore, from the viewpoint of the technical solution, the organic solvent virus inactivating agent in the present invention is distinguished from the organic solvent in the general sense.
  • the organic solvent virus inactivating agent includes any solvent for virus inactivation, such as tributyl phosphate (abbreviated as TnBP in the present invention), formaldehyde, diethyl ether, and the like. It is worth noting that organic solvent virus inactivating agents are often shared with detergents.
  • the detergent includes any detergent for virus inactivation, such as a Tween-type detergent (e.g., Tween-80) and a Triton-based detergent (e.g., Triton X 100).
  • the photosensitizer virus inactivation comprises virus inactivation with a photosensitizer virus inactivating agent as a virus inactivating agent; the term "photosensitizer virus inactivating agent" means under specific conditions of virus inactivation treatment.
  • reaction kinetic conditions are less favorable (eg, liquid
  • concentration of the virus inactivating agent when the phase virus is inactivated is generally not more than ⁇ /l
  • the reaction conditions are relatively conservative (for example, usually the reaction temperature is less than 38* ⁇ and 11 is between 6-10);
  • the photosensitizer virus inactivating agent in the present invention is distinguished from the photosensitizer in the general sense.
  • photosensitizer virus inactivating agents include, but are not limited to, dye photosensitizer virus inactivating agents, hypericin and psoralen photosensitizer virus inactivating agents, and the like. Generally, these photosensitizer virus inactivating agents can have much higher virus inactivation efficiencies under appropriate light exposure.
  • the term "dye-based photosensitizer virus inactivating agent” means a dye-like substance which can be used as a photosensitizer virus inactivating agent for virus inactivation (for example, photochemical virus inactivation), for example, phthalocyanine (US Patent No.
  • Psoralen-type photosensitizer virus inactivating agent means that the photosensitizer virus inactivating agent can be used for virus inactivation (eg photochemistry)
  • Viral inactivated psoralen substances U.S. Patents 4,169,204, 4,294,822, 4,328,239 and 4,727,027), including psoralen (English Psoralen ⁇ psoralen derivatives, etc. Examples of psoralen derivatives)
  • psoralen Korean Psoralen ⁇ psoralen derivatives, etc. Examples of psoralen derivatives
  • side effects refers to the ability of a solid phase medium, treatment system or device to have unwanted functions in addition to having a useful function, and may also cause an undesirable change in the biological activity of the biological fluid, such as the following One or more changes: a reduction in the total amount of protein, interference with the coagulation system, changes in platelet morphology, improvement in hemolytic properties, and the like.
  • the term "specific adsorption” refers to adsorption of a virus inactivating agent to a virus inactivating agent, or removal of leukocyte media by leukocyte medium;
  • non-specific adsorption refers to a virus inactivating agent adsorption medium pair Adsorption of substances other than viral inactivating agents, or removal of substances other than leukocytes by leukocyte mediators.
  • Non-specific adsorption includes adsorption (e.g., degreasing) and unwanted adsorption (e.g., reduction in clotting factor activity) that are beneficial to the desired properties of the biological fluid.
  • Unwanted reactivity of the solid phase medium includes unwanted adsorption and unwanted other reactive activities (e.g., catalytic activity, enzyme activating activity, activity involved in reactions in biological fluids, etc.).
  • solid phase medium also referred to as medium, refers to a solid phase material having a certain function (for example, adsorption function, virus inactivation function, etc.);
  • virus inactivating agent adsorption medium Sometimes referred to as an adsorption medium, it means having at least, but not limited to, a function of adsorbing a virus inactivating agent or/and a virus inactivating agent derivative (for example, sometimes it may also have a filtering function, a leukocyte function, etc.), And can ultimately reduce the concentration of viral inactivating agents or/and virus inactivating agent derivatives in biological fluids to acceptable levels (eg, tributyl phosphate concentration less than 10 ppm, methylene blue concentration less than 0.03 g / ml, etc.
  • acceptable levels eg, tributyl phosphate concentration less than 10 ppm, methylene blue concentration less than 0.03 g / ml, etc.
  • the solid phase material, the adsorption medium may be a pure substance (such as activated carbon powder, wood fiber, etc.), or may be a mixture (for example, a cellulose-containing filter medium, an activated carbon-containing filter medium, a C18 chromatography gel, etc.)
  • the term "viral inactivating medium” means a solid phase medium having at least, but not necessarily limited to, a virus inactivating function (for example, it may also have a filtering function sometimes), such as a virus inactivating agent/adsorption medium complex, the term" Interleukin "refers to any solid phase media may be used to leukocytes.
  • the term "viral inactivating agent adsorbent” means an adsorbent which is effective for removing a virus inactivating agent by adsorption under specific conditions of virus inactivation treatment. These specific conditions include: specific viral inactivating agents; specific adsorption environments (eg, biological fluids often contain components that affect adsorption, such as lipids in human plasma); specific adsorption kinetic conditions (eg, eg In the embodiment of the present invention, the concentration of the virus inactivating agent in the biological liquid is small, but the concentration of the virus inactivating agent remaining after the adsorption is required to be extremely low, for example, less than 10 ppm or less than 0.01 ⁇ 1 / 1) ; specific adsorption reaction conditions ( For example, to maintain the activity of the biological fluid, the adsorption temperature is usually limited to 0-37 ° C, the adsorption pH is usually limited to 3.8-9.8); Thus, from the viewpoint of the technical solution, the virus inactivating agent a
  • organic polymer chemical adsorbent is prepared by introducing a sufficient amount of a bonding group (for example, an ionic group, a coordinating group, etc.) which generates a chemical bond to an organic polymer, thereby Mainly through the introduced bonding groups to generate chemical bonds (such as ionic bonds, coordination bonds, etc.) into the reagents, such as ion exchange fibers, ion exchange particles and chelating agents;
  • "organic ⁇ molecular affinity adsorption "agent” is prepared by introducing a sufficient amount of an affinity ligand (eg, a terminal group of hydrazine, or an affinity of a viral RNA or DNA for the affinity adsorption of certain photosensitizer virus inactivating agents) onto an organic hydrazine molecule, Therefore, the reagent mainly adsorbed by the introduced affinity ligand, such as affinity fiber, affinity particle, etc.; "organic polymer physical adsorbent” refers to
  • the organic polymer does not include activated carbon.
  • the organic polymer physical adsorbent does not include the currently known organic polymer affinity adsorbent and the organic polymer ion exchange adsorbent, and these functionalized organic polymers are subjected to special affinity grouping ( Modifications such as affinity fibers, affinity particles) or/and specialized ion exchange groups (eg, ion exchange fibers, ion exchange particles).
  • the organic polymer physical adsorbent does not have to be prepared by introducing a sufficient amount of ion exchange groups or organic ligands onto the organic polymer, and thus mainly through weak interactions on the organic polymer (including van der Waals attraction, dipole-dipole Interaction, hydrogenation, etc.) and adsorption of a small amount of (end group, or / and 5% or less substituents on the chain) ionic groups (for example, a small amount of acid groups on plant fibers) introduced during the preparation process Reagents.
  • the organic polymer in the term "organic polymer physical adsorbent" includes a natural polymer, such as a strong natural polysaccharide.
  • the polysaccharide comprises one or more of the following: cellulose, dextran, agarose, chitosan, starch.
  • the cellulose derivative include nitrocellulose, cellulose acetate, methyl cellulose, carboxymethyl cellulose, and the like.
  • polyglycans have been used as carriers in addition to viral inactivating agents, for example, nitrocellulose has been used as a carrier for photosensitizer virus inactivating agent ligands in addition to photosensitizer virus inactivating agents and leukocyte-removing filters (US patent) US08/179567, 08/204102, 08/347564), a polysaccharide containing polypolysaccharide is used as a carrier for an SD agent adsorbent (for example, C18) for use in addition to an SD agent, etc., another meaning of the embodiment of the present invention
  • an SD agent adsorbent for example, C18
  • certain filters e.g., Seitz-BiolO, Seitz-Bio20, Seitz-bio 40
  • certain porous particles e.g., Sephadex G10, Sephadex G25 Sephadex G50, Sepharose
  • the organic polymer in the term "organic polymer physical adsorbent" includes synthetic polymers such as polyolefins, polyurethanes, polydecyl esters, and the like.
  • the synthetic polymer includes a polyolefin such as polystyrene, polyvinyl chloride, polyvinyl alcohol, or the like.
  • certain filters eg Wurz-Supradurl00, Seitz-Supradur200, Seitz-Supradur 500
  • certain Porous particles for example, crosslinked acrylic macroporous adsorption resin, crosslinked polystyrene macroporous adsorption resin, polystyrene-polyacrylonitrile macroporous resin, polymethyl methacrylate resin, polyisobutylene, etc.
  • the term "passivating agent” refers to a reagent which has a passivation effect on an undesired reaction (eg, side effects) on a solid phase medium or other composition of a treatment system, but may not be limited to passivation.
  • the term “composite” means a material in which two or more compositions are joined by adsorption or other physicochemical, chemical, or biochemical interaction.
  • the term "passivation agent/adsorption medium complex” refers to a complex of at least an adsorbent medium composed of a virus inactivating agent and a passivating agent of an adsorbent medium;
  • the term "viral inactivating agent / adsorbent medium / passivator complex” means a minimum composition of a virus inactivating agent a complex of an adsorption medium for the virus inactivating agent and a passivating agent for the adsorption medium.
  • the processing system can be used as a device (eg, a filter, a chromatography column, etc.), or as a component of the device (eg, a filter, column, etc. in a single blood or component processing device). Wait).
  • the term "device” means a structure in which the minimum composition is the above-described treatment system, which can be practically applied to the treatment of biological fluids, such as filters, chromatography columns, kits (English Kit:), etc.; "Case” means a hollow vessel that can hold at least a solid phase medium, such as a hollow filter cartridge of a filter, a hollow column of a chromatography column, and the like.
  • the term "filter material” generally means a solid phase medium which can only allow a structure below a certain size to flow. Therefore, the filter medium in the present invention is often used as a stationary phase in a solid-liquid phase reaction, for example, a filter material having an adsorption function for a virus inactivating agent (referred to as a specific adsorption filter medium). Whether a filter material specifically adsorbs the filter material, in addition to the information provided by the manufacturer (such as activated carbon filter plate), it is mainly to see if it has a specific adsorption function in the virus inactivation treatment.
  • a Seitz-Bio filter plate which is generally considered to have no adsorption function is found to be a dye-based photosensitizer virus inactivating agent adsorption medium in the examples of the present invention.
  • the filter media includes absolute filter media and depth filter media.
  • a preferred embodiment of the filter material of the present invention is a depth filtration filter.
  • the fluid flow path in the deep filter media is long and has a significant impact on its function (eg adsorption, virus inactivation).
  • Filter media come in many forms, such as filters, filter plates, filter rods, filter cartridges, filter mats, and more.
  • the term "filter” means a device comprising at least a filter material and its holder, including but not limited to known filters.
  • the holder includes a structure that seals the periphery of the solid phase medium with pressure, such as a filter disc, a filter cartridge, a filter column, and the like.
  • the holder generally includes: a container for accommodating the solid phase medium, an upper end structure and a lower end structure for fixing the solid phase medium, and a pressure sealing filter for surrounding the solid phase medium
  • the structure of the solid-state medium of the material for example, a pressure-closed structure in which the solid phase medium is closed to avoid liquid leakage due to the contact force of the convex body which is higher on the upper end structure and the lower end structure to the periphery of the solid phase medium when closed).
  • other functional structures can be added, such as in the upper structure.
  • a structure with a special function introduced on the lower end structure for example, a sterile filter membrane, etc.
  • the term "dyeing” means a structure which can bind a dye, including a physical structure, a physicochemical structure, a chemical structure (for example, a group), and the like;
  • the term “fiber” means a length/diameter ratio of more than 10, and A material having an average diameter of less than 1 mm;
  • the term “Fibrets” refers to a fiber having a large number of irregular micro branches and a large surface area formed by a special production process or special processing. Structure, this fiber microstructure is very short (less than 1 mm) and very fine (less than 50 ⁇ m), for example Seitz-BiolO, Seitz-Bio20, Seitz-bio40 filter plates contain fiber microstructure.
  • a first aspect of the invention relates to a virus inactivation method of the invention.
  • the virus inactivating method of the present invention comprises at least: A) providing a biological fluid; B) providing a treatment system; and C) contacting the biological fluid with the treatment system, wherein the treatment system comprises at least a solid phase a medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, the virus inactivating medium comprising a virus a living agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: 0) a passivation adsorption medium, the passivation adsorption medium containing at least an adsorption medium and being fixed on the adsorption medium a passivating agent; or/and (b) a physical adsorbing medium comprising: (i) a photosensitizer physic adsorbing fiber used as a photosensit
  • the permissible residual value of the virus inactivating agent in the bio-liquid product standard is usually very low (for example, the TnBP approved residue value in the European Pharmacopoeia is less than 10 ppm)
  • the general view is: It is not suitable for passivation of the adsorption medium.
  • the adsorption medium can still be used for adsorption removal of the virus inactivating agent, and has less side effects than the adsorption medium.
  • the general view is that the virus is inactivated medium (especially the solid phase of the virus inactivating agent not on the long-chain adsorption group) Passivation of a medium, such as SD-activated carbon, may reduce the inactivation efficiency of the virus inactivating medium. Thus, no passivation treatment was introduced in the development of the virus inactivating medium.
  • embodiments of the present invention will demonstrate that the use of a virus inactivating medium containing a passivating agent not only effectively protects the biological activity of the liquid medium contacted by the solid phase medium, but also effectively inactivates the virus, thereby achieving The object of the invention.
  • the leukocyte-removing medium usually has more or less adsorption capacity to white blood cells.
  • the general idea is that the addition of a passivating agent can inhibit its side effects, but it may also reduce its adsorption on white blood cells. Surprisingly, however, in one embodiment of the invention, the addition of a passivating agent inhibits its side effects, but it can still be used to effectively remove leukocytes.
  • the first treatment system of the invention can be used to simultaneously remove viral inactivating agents (or/and viral inactivating agent derivatives) and white blood cells.
  • the removal of additives by solid phase media containing adsorbents is a well-known technique that has long been used.
  • the viral inactivating agent can be removed by a solid phase medium containing a viral inactivating agent adsorbent. Since the solid phase medium with adsorption function is generally very much, and the virus inactivating agent includes many kinds, the virus inactivating agent adsorption medium containing the adsorption function for the specific virus inactivating agent is developed.
  • Certain processing systems have been a very difficult creative task, and it has become more difficult to develop a treatment system that has a specific adsorption of a specific viral inactivating agent without substantial non-specific adsorption (for example, essentially no side effects:). Creative work.
  • photosensitizers for physically adsorbing fibers is also based on the unexpected results of embodiments of the present invention.
  • certain dyes eg, methylene blue
  • fibers containing dyed seats eg, natural fibers
  • C Even filter media containing natural fibers have long been used for the production of biological products, but no adsorption medium for directly using natural fibers as a dye-based photosensitizer virus inactivating agent has been seen so far.
  • these fibers are not only adsorbable to the dye-based photosensitizer virus inactivating agent, but also have the characteristic that the adsorption is sufficiently strong (for example, the concentration of methylene blue after adsorption is reduced to 0.03 g / Mol is below); its adsorption is kinetically stable (for example, there is no significant desorption of methylene blue adsorbed on the filter plate when the biological liquid is continuously flowing); its adsorption is reproducible (for example, under the same conditions) The adsorption reproducibility is 100%); and its adsorption is sufficiently specific (for example, substantially no side effects).
  • the adsorption is sufficiently strong (for example, the concentration of methylene blue after adsorption is reduced to 0.03 g / Mol is below); its adsorption is kinetically stable (for example, there is no significant desorption of methylene blue adsorbed on the filter plate when the biological liquid is continuously flowing); its adsorption is reproducible (for example, under the same conditions) The a
  • the present invention is not intended to be discussed in theory, and only some of the fibers containing organic polymeric physical adsorbents are provided for some observations. Perhaps due to the following reasons, weak interactions of organic polymeric physical adsorbents (such as trace ionic groups introduced during production, van der Waals attraction, dipole-dipole interaction, hydrogen chemistry) are strong enough to effectively adsorb dyes Photosensitizer virus inactivating agent: 1). Dye-based photosensitizer The "dye up-take" of the virus inactivating agent is usually a biological liquid containing a fiber as a stationary phase and a dye-based photosensitizer virus inactivating agent.
  • the phase is carried out under the conditions of the preferred solid-liquid phase reaction; 2) the dye-based photosensitizer virus inactivating agent is not required to be uniformly dyed; 3) the dye-based photosensitizer virus to be adsorbed in terms of the amount of fibers used
  • the total amount of inactivating agent is not very large; 4 ).
  • organic solvent physical adsorption media is also based on the unexpected results of embodiments of the present invention.
  • an organic solvent virus inactivating agent eg TnBP
  • porous particles containing polyolefin can remove oil stains
  • the inactivating agent is combined to prepare a virus inactivating medium.
  • the porous particles not only have an adsorption capacity to the organic solvent virus inactivating agent, but also have the characteristic that: the adsorption is sufficiently strong (for example, the concentration of TnBP after adsorption is reduced to less than 10 ppm); The adsorption is kinetically stable (eg, no significant TnBP desorption occurs); its adsorption is reproducible (eg, its adsorption reproducibility is 100% under the same conditions); and its adsorption is sufficiently specific (eg , basically no side effects).
  • the solid phase medium comprises an organic solvent virus inactivating agent adsorption medium
  • the organic solvent virus inactivating agent adsorption medium contains at least the passivation adsorption medium. Or / and organic solvent physical adsorption medium.
  • the solid phase medium comprises an organic solvent virus inactivating medium, and the organic solvent virus inactivating medium contains at least an organic solvent virus inactivating agent, and the blunt The adsorption medium, or/and the organic solvent physical adsorption medium.
  • the organic solvent physical adsorption medium comprises a macroporous adsorption resin or/and a fiber.
  • the fiber contains an organic polymer physical adsorbent containing an organic solvent virus inactivating agent, such as natural fiber (for example, Seitz Bio series filter plate), polyolefin fiber (for example, Seitz). Supradur 80, Seitz Supradur500, Seitz Eco 1000, etc.), etc.
  • the macroporous adsorption resin (organic solvent physical adsorption resin) used includes polystyrene particles (for example, Amberlite XAD-7HP, Amberlite AD-16), phenolic polycondensation type particles, polyacrylate particles, and polyalkyl ester-containing macroporous adsorption resin.
  • the organic solvent physical adsorption medium further includes a porous non-particulate material having an inner surface containing the organic polymer. For example, polyolefin coated ceramics, and many more.
  • the organic polymer physical adsorbent of the organic solvent virus inactivating agent comprises a polyolefin.
  • the polyolefin comprises polystyrene, especially a macroporous adsorption resin comprising polystyrene.
  • the solid phase medium comprises a photosensitizer virus inactivating agent adsorption medium
  • the photosensitizer virus inactivating agent adsorption medium contains at least the passivation adsorption medium. Or / and the photosensitizer physically adsorbs the fibers.
  • the photosensitizer physical adsorbing fibers used include natural fibers, cellulose-based derived fibers, and non-functionalized synthetic fibers.
  • the photosensitizer physically adsorbing fibers comprise natural fibers.
  • the natural fibers used include plant fibers, animal fibers (e.g., silk).
  • the plant fibers used include cotton fibers (e.g., non-fat cotton), wood fibers (in filter plates), hemp fibers, pulp (e.g., paper).
  • the fibers also include cellulose based derived fibers (e.g., viscose, acetate, and the like).
  • the natural fibers comprise wood fibers.
  • the photosensitizer physically adsorbing fibers comprise synthetic fibers, preferably non-functionalized synthetic fibers.
  • the non-functionalized synthetic fibers used include synthetic fibers other than affinity fibers and ion exchange fibers, such as polyester fibers, polypropylene fibers (eg, ultrafine polypropylene oil absorbing fibers), polyamide fibers, Polyurethane fibers and other polyolefin fibers.
  • the fiber has one or more of the following characteristics: A). Average diameter 0.01-20 ⁇ ; ⁇ ). Curl degree 1-2, ⁇ C). Water absorption 25-50 g /100 g fiber.
  • other structural parameters of the fiber such as the glass transition temperature of the polymer, the electrokinic layer potential of the fiber, and the isoelectric point, etc., also affect its adsorption capacity.
  • the synthetic fiber comprises a polyolefin fiber.
  • certain filter media e.g., Seitz-SupradurlOO, Seitz-Supradur 200 Seitz-Supradur 500
  • a sensitizing agent for the photosensitizer contain polyolefin fibers.
  • the virus inactivating agent adsorption medium further comprises particles, and the particles comprise an organic polymer physical adsorbent of a photosensitizer virus inactivating agent.
  • the photosensitizer virus inactivating agent adsorption medium comprises styrene-containing organic polymer particles and a non-woven fabric or filter of photosensitizer physico-adsorbing fibers (for example, polyolefin fibers) located under the particles. material.
  • the concentration of the photosensitizer virus inactivating agent when the virus is inactivated is from 0.55 to 3.0 ⁇ ⁇ 1 /1.
  • a high photosensitizer virus inactivating agent concentration is beneficial to reduce the time to virus elimination, or / and to apply virus-eliminating conditions that are beneficial to maintain biological activity (eg lower temperature) Degree, lower light intensity, etc.).
  • the side effect of a solid phase medium containing a viral inactivating agent adsorption medium is one factor limiting the concentration of the photosensitizer virus inactivating agent.
  • the virus inactivation is carried out at a temperature below room temperature, for example between 4-19.5 ° C, preferably between 15 and 19.5 ° C, more preferably between 15 and 17 ° C. Row.
  • the solid phase medium comprises the organic solvent virus inactivating medium, an organic solvent virus inactivating agent adsorption medium, or/and a photosensitizer physical adsorption fiber. This is a dual virus inactivation method.
  • the solid phase medium comprises an iodine virus inactivating medium, the iodine virus inactivating medium comprising at least iodine, an iodine adsorption medium, and a passivating agent.
  • the solid phase medium comprises a solid phase virus inactivating agent and a passivating agent immobilized thereon.
  • a solid phase virus inactivating agent is a positive charge deep filtration filter (e.g., Cuno Corporation Zetaplus VR filter plate).
  • the solid phase virus inactivating agent is a solid phase virus inactivating agent based on a positive charge action.
  • the virus inactivating medium has a height greater than 2 cm and less than 6 cm, and the virus inactivating agent adsorption medium has a height greater than 1 cm and less than 3 cm.
  • the virus inactivating agent adsorption medium has a height greater than 1 cm and less than 3 cm.
  • the solid phase medium comprises a depth filter medium, the deep filter medium containing the virus inactivating agent adsorption medium, and having the following characteristics: A) The average density is greater than 0.25 g/cm 3 ; and B). The ash content is less than 1%. According to an embodiment of the invention, controlling ash has important implications for minimizing side effects.
  • the filter medium may contain, in addition to the fibers, an adhesive, a reinforcing agent (e.g., polyolefin), and the like. Higher density filter media can have higher adsorption capacity.
  • the solid phase medium comprises one of the following filters or their analogs: Seitz-BiolO, Seitz-Bio20, Seitz-bio40, Seitz-SupradurlOO, Seitz-Supradur200, Seitz-Supradur500, Seitz -Supradurl000.
  • the average density of the filter media is greater than
  • the depth of the depth filter media is greater than 6 mm.
  • the passivating adsorption medium may contain one or more passivating agents, one or more adsorbing media, and the like.
  • the adsorption medium in the passivation adsorption medium includes: a virus inactivating agent adsorption medium, a virus inactivating agent adsorption medium in the virus inactivating medium, and a leukocyte removal medium.
  • the virus inactivating agent adsorption medium contains one or more of the following adsorbents: activated carbon, inorganic adsorbent, Organic polymer chemical adsorbent, organic polymer affinity adsorbent, organic germanium molecular physical adsorbent.
  • the organic polymer affinity adsorbent includes particles or fibers to which a ligand (for example, a C6-C18 carbon chain, a hydrazine-containing end group or a viral RNA, or a DNA replica) is immobilized. These adsorbents have an adsorption function for virus inactivating agents, but tend to have strong undesired reactivity.
  • a ligand for example, a C6-C18 carbon chain, a hydrazine-containing end group or a viral RNA, or a DNA replica
  • the adsorbing medium in the passivating adsorption medium comprises activated carbon.
  • activated carbon occurs in various forms such as activated carbon powder, activated carbon felt, activated carbon-containing filter material, and the like.
  • the adsorbing medium in the passivating adsorption medium comprises an ion exchange adsorption medium.
  • the ion exchange adsorption medium includes: an inorganic adsorbent such as diatomaceous earth, perlite, glass fiber, etc.; an organic polymer chemical adsorbent such as an ion exchange resin, an ion exchange fiber, or the like.
  • the adsorption medium in the passivation adsorption medium comprises the above physical adsorption medium (organic solvent virus inactivating agent adsorption medium, photosensitizer physical adsorption fiber). Passivation of the physical adsorption medium makes it possible to further reduce side effects.
  • the passivating agent is preferably an organic substance that can be injected into a human body.
  • These passivating agents are characterized by: 1) themselves being relatively inert; 2) can be immobilized on a solid phase medium to inhibit its side effects without meaningful adverse effects on its function; 3).
  • the controlled amount of passivating agent is detached from the solid phase medium or other composition into the biological fluid, which does not cause substantial damage to the safety of use of the biological fluid.
  • the passivating agent in the passivation adsorption medium comprises a hydrophilic group or/and a lipophilic group, and the solid phase medium pair can be lowered
  • An organic substance that describes the side effects of biological fluids An organic substance having an oleophilic group is poorly water-soluble (for example, a natural oil, an organic solvent virus inactivating agent), and a considerable number of organic substances having a hydrophilic group or having a hydrophilic group and a lipophilic group are considered to have poor adsorption force ( For example, surfactants, hydroxy compounds, amino acids), no passivating agents are used as solid phase media. Surprisingly, such materials are useful as passivating agents in the practice of the present invention and are intended to serve the objectives of the present invention.
  • the passivating agent content in the solid phase medium is greater than It is preferably greater than 1 ⁇ 1/ ⁇ 3 .
  • the purpose of the present invention e.g., minimizing side effects
  • the adsorption of viral inactivating agents e.g., methylene blue
  • the amount of certain passivating agents on the activated carbon material is greater than 5 ⁇ 1/ ⁇ 3 .
  • the upper limit of the content of the passivating agent it should be different according to its activity (specific adsorption capacity or virus inactivation ability) and the degree of passivation required (for example, the degree of passivation of biological fluids with or without clotting factors may be different) , optimized according to known methods.
  • the purifying agent comprises a natural fat.
  • Place Natural oils and fats include natural oils and natural oils and fats. These materials all have an oleophilic group.
  • the natural fats and oils include vegetable oils and phospholipids, wherein: vegetable oils include castor oil, soybean oil, tea oil, and glycerin; and phospholipids include cephalin, lecithin, and plant phospholipids.
  • Examples of natural oil and fat derivatives include vegetable oil emulsions, fat emulsions, and the like.
  • the purification agent comprises a hydroxy compound.
  • the hydroxy compound has a hydrophilic group including alcohols (for example, ethanol, glycerin, glycerin sodium chloride, and the like), saccharides, or/and their respective derivatives.
  • the saccharide comprises a monosaccharide, a polysaccharide, or/and a polypolysaccharide, for example: glucose, mannitol (Molecular Formula C 6 H 14 0 6 ), Sorbitol, Glycerol Fructose, Sucrose, Hydroxyethyl Starch, lentinan, etc.
  • the purification agent comprises an amino acid.
  • the amino acid has a hydrophilic group including cystine, lysine, tyrosine, glycine, arginine, valine, serine, a polymer of these amino acids, and the like.
  • the surprising result of embodiments of the present invention is the saccharide or /amino acid and can be adsorbed on the surface of the activated carbon and reduce the side effect of the activated carbon on the biological fluid. Polyamino acids also have a similar effect.
  • the purification agent comprises an organic solvent.
  • the organic solvent includes an organic solvent virus inactivating agent such as tributyl phosphate (TnBP), diethyl ether, glycerin, ethyl carbonate, ethyl lactate, benzyl benzoate, and the like.
  • TnBP tributyl phosphate
  • diethyl ether diethyl ether
  • glycerin glycerin
  • ethyl carbonate ethyl lactate
  • benzyl benzoate benzyl benzoate
  • the passivating agent comprises a polypeptide.
  • the polypeptide has hydrophilic groups such as albumin, serum, and milk, and their respective derivatives.
  • the passivating agent comprises a surfactant.
  • the surfactant has a hydrophilic group and a lipophilic group.
  • the present inventors have found that a large number of surfactants, especially surfactants having both hydrophilic groups and lipophilic groups in the molecular structure, can be adsorbed on the surface of activated carbon and reduce the side effects of activated carbon on biological fluids.
  • Hydrophilic groups include: -OH, -OS0 3 , - (C3 ⁇ 4CH 2 0) 3 , -N(CH 3 ) 2 , -N(CH 3 ) 3 , -CH 2 COO, -NH 2 ; lipophilic groups Including: organic ring groups, -(CH 2 i-, etc.
  • surfactants include: Triton-based surface active substances, Tween-based surface active substances, sodium cholate, tetrahydrofurfural polyglycol ether, two Methylacetamide, polyvinylpyrrolidone, and dimethyl sulfoxide. Examples of partial surfactants as passivating agents are given in the examples.
  • the virus inactivating medium is immobilized with an organic solvent virus inactivating agent and an organic solvent deactivator, and the organic solvent deactivator and the organic solvent virus are inactivated.
  • the total content of the agent is greater than 0.2 ⁇ 1/ ⁇ 3 , preferably greater than 0.3 ⁇ 1/ ⁇ 3 .
  • the examples of the present invention show that only when the content of the organic solvent virus inactivating agent (for example, ⁇ ) is greater than a certain degree, the organic solvent can be used as a virus inactivating agent for effective virus inactivation and as a passivating agent. Minimize side effects.
  • the embodiment of the present invention demonstrates that the content of the organic solvent virus inactivating agent is greater than 0.2 mol/cm 3 .
  • the solid phase medium may also contain an appropriate amount of detergent D (e.g., greater than 0.01 mol/cm 3 ) or other passivating agent (e.g., polysaccharide).
  • D e.g., greater than 0.01 mol/cm 3
  • other passivating agent e.g., polysaccharide
  • the passivating agent comprises a combination of two or more of the above passivating agents, for example: dextran 40 / glucose, dextran 40 / albumin, albumin / glucose, TnBP / spit Wen-80/glucan, and so on.
  • the treatment system further comprises a leukocyte-free solid phase medium.
  • an embodiment of the method of the present invention comprises at least: A) providing a single blood component; B) providing said treatment system, said treatment system further comprising a leukocyte solid phase medium; C). Flowing the biological fluid through the treatment system for leukocyte removal and virus inactivation or/and viral inactivating agent adsorption; D). The treatment system from C) is substantially free of the virus inactivating agent and A single blood component of white blood cells.
  • the leukocyte-removing solid phase medium comprises the organic polymeric physical adsorbent.
  • the leukocyte-removing medium medium comprises the virus inactivating agent adsorption medium.
  • the biological fluid comprises a single human blood component.
  • the content of the organic solvent virus inactivating agent in the organic solvent virus inactivating medium is equal to or less than 3 X (10 ppm of the volume of the single blood component) Ml), or less than (lOppmX the volume of the single-part blood component ml). In fact, limiting the amount of organic solvent virus inactivating agent may help to eliminate the detection of organic solvent virus inactivating agents in the final product.
  • a treatment system for effectively reducing virus hazards of the present invention is the treatment system described in the above virus inactivation method.
  • the device includes a column, a filter, a kit, and the like.
  • some of the devices further comprise a viral inactivating agent addition structure, or/and a virus inactivation reaction site, or/and a wash solution.
  • the device is a single human blood component treatment kit.
  • the activated media for the activated carbon-containing virus inactivating agent are activated carbon powder, activated carbon felt (ZC-1200A, China Zichuan Carbon Fiber Co., Ltd.) and activated carbon filter plates (AKS 5 and AS 6, respectively). Seitz, Germany).
  • the virus inactivating agent adsorption medium containing the ion exchange adsorbent is silicon oxide particles and glass fibers, respectively.
  • the material containing the silicon oxide particles was a perlite-containing filter plate (Zetaplus Delipid, Cuno Corporation).
  • the virus inactivating agent adsorption medium containing the affinity adsorbent is C18 reverse phase gel (Water Company, USA).
  • Physical adsorption medium Among them, the physical adsorption media used are:
  • Photosensitive agents Physically adsorbed fibers, including natural fibers, cellulose-based derived fibers, and non-functionalized synthetic fibers.
  • the natural fibers used include plant fibers, animal fibers (e.g., silk).
  • the plant fibers used include cotton fibers (e.g., non-fat cotton), wood fibers (in filter plates such as Seitz Bio series filter plates), and pulp (e.g., paper).
  • the non-functionalized synthetic fibers used include synthetic fibers other than affinity fibers and ion exchange fibers, such as polyester fibers, polypropylene fibers (for example, ultrafine polypropylene oil absorbing fibers), polyamide fibers, and polyurethane fibers (both non-woven fabrics). ).
  • polyolefin fibers are contained in the filter material used as the sensitizer physical adsorption medium in the following examples, such as Seitz Supradur 80, Seitz Supradur 500 Seitz Eco 1000, and the like.
  • Organic solvent physical adsorption medium including fibers (organic solvent physical adsorption fibers) and macroporous adsorption resins (organic solvent physical adsorption resins).
  • the fiber used contains an organic polymer physical adsorbent containing an organic solvent virus inactivating agent, such as natural fibers (for example, Seitz Bio series filter plates), polyolefin fibers (such as Seitz Supradur 80, Seitz Supradur 500, Seitz Eco 1000, etc.), and many more.
  • the macroporous adsorption resin (organic solvent physical adsorption resin) used includes polystyrene particles (for example, Amberlite XAD-7HP, Amberlite XAD-16), phenolic polycondensation type particles, polyacrylate particles, and polyalkyl ester-containing macroporous adsorption resin.
  • Amberlite XAD- 7HP is a polar polystyrene macroporous resin particle
  • Amberlite XAD-16 is a non-polar polystyrene macroporous resin particle.
  • the particles used have one or more of the following characteristics: A). Average particle size 5-1000 ⁇ ; ⁇ ). Specific surface area 100-2000 m dry powder; C). Average pore diameter less than 5-600 A; D) Excluding volume less than 50,000 molecular weight .
  • the porous protein solid phase medium used in part has a spherical protein exclusion lower limit molecular weight of less than 5,000.
  • the photosensitizer physical adsorbing fiber or the organic solvent physically adsorbing fiber comprises a fiber filament, a nonwoven fabric, a cloth, and a depth filter medium.
  • the prefix Seitz- refers to the product of the German company Sdtz.
  • the permeability of some of the deep filter media used in the solid phase media is less than 200 L/m 2 min, which are: Seitz-Supradur 100, Seitz-biolO, Seitz-bio 40 0
  • the passivating agent used includes:
  • a natural fat or oil having an oleophilic group or/and an organic solvent for example, castor oil, soybean oil, tea oil, fat milk, tributyl phosphate (abbreviated as ⁇ in the present invention), and diethyl ether.
  • Surfactants having a hydrophilic group and an oleophilic group for example: sodium cholate, Triton® 100, Tween 80, and polyvinylpyrrolidone.
  • Hydrocarbyl compounds having a hydrophilic group such as: glycerin, glucose, maltose, dextran (dextran), water-soluble cellulose, mannitol (C 6 H 14 0 6 ), sorbitol, hydroxyethyl Starch, lentinan.
  • Amino acids having a hydrophilic group such as: valine, arginine and glycine.
  • Polypeptides with hydrophilic groups such as: human albumin (Beijing Tiantan Biological Products Co., Ltd.), Buxuekang (Germany Biotest) and milk.
  • a composite passivating agent for the above passivating agent for example: dextran 40/glucose, dextran 40/albumin, albumin/glucose, TnBP/Tween-80/dextran.
  • dextran 40/glucose for example: dextran 40/glucose, dextran 40/albumin, albumin/glucose, TnBP/Tween-80/dextran.
  • virus inactivating agent used includes:
  • Organic solvent virus inactivating agent or organic solvent virus inactivating agent/detergent for example. ⁇ Tributyl phosphate, diethyl ether, TnBP/Tween-80, TnBP/Triton X100, TnBP/sodium cholate, diethyl ether /Tween 80.
  • Photosensitizer virus inactivating agents such as dye virus inactivating agents or psoralen virus inactivating agents.
  • the dye-like virus inactivating agents used were methylene blue, methyl violet, and toluidine blue, respectively.
  • the psoralen virus inactivating agents used were psoralen and 8-m e th OX ypr OT al en , respectively . It should be understood by the skilled person that other psoralens have almost the same adsorption properties as the psoralen used.
  • Solid phase virus inactivating agents such as Zetaplus VR filter (Cuno:).
  • the conditions for virus inactivation are generally as follows: pH is 2.0-12.0; temperature is -5 to 6 CTC; amount of solid phase medium and biological liquid containing virus inactivating agent according to virus inactivation experiment Preferably (for example, solid medium volume / biological liquid volume between 1% and 30%); virus inactivation hydrodynamic conditions are preferred according to virus inactivation experiments (eg linear flow rate 0.1-lOcm/min, pressure 0.1-) 5 kg/cm 2 ); the amount of solid phase medium and biological liquid containing the virus inactivating agent adsorption medium is preferably according to the virus inactivating agent removal experiment (for example, the solid medium volume/bioliquid volume is between 1% and 30%) Between) The determination of the liquid phase adsorption reaction conditions is preferably carried out in accordance with a well-known rule, for example: the greater the flow rate of the biological liquid, the smaller the adsorption efficiency;
  • the virus inactivation efficiency was carried out according to a known method using a pattern lipid enveloped virus.
  • VSV initial titer greater than 10 4 ⁇ 8
  • Sendai virus initial titer greater than 10 5 ⁇ 1
  • the detection of various biological activities was carried out by a known method.
  • the leukocyte removal rate is measured and calculated by a known method.
  • the devices prepared in the following examples include devices that can be used to effectively reduce the pathogen hazard of a single human blood component. It comprises at least the treatment system of the invention, wherein the chamber is a conventional filter cartridge or chromatography column.
  • the solid phase media loading height is 3-5cm, and there is a 0.4 ⁇ filter at the outlet end of the filter to prevent the media debris from leaking out.
  • the column prepared by the following examples is effective for reducing the pathogen hazard of a single blood component, generally having an inner diameter of 1 cm, a solid phase medium loading height of 3-5 cm, and a 0.4 ⁇ m filter at the outlet end of the column. In case the media debris leaks out.
  • Example 1.1 Virus inactivating agent adsorption medium containing passivating agent
  • the solid phase medium prepared in this example is an adsorption medium/passivator composite.
  • the passivating agent is at a preferred concentration (for example, natural fat: 0.2-0.5%; organic solvent: 0.3-2.0%; surfactant: 1.0-3.0%; polypeptide: 1.0-10.0%; amino acid: 3.0 -10.0%; etc.) Disperse in an aqueous solution (eg PBS buffer).
  • aqueous solution eg PBS buffer
  • other media e.g., organic solvent virus inactivating agents
  • a passivating agent dispersion medium e.g., solution, suspension
  • concentration of the dispersion treatment system w/v
  • the concentration of the dispersion treatment system may be from 0.1% to 50°/. between.
  • the dispersion of the low water-soluble organic substance deactivator can be carried out by a known technique. There are many methods for dispersing natural fats and oils in aqueous solutions, such as emulsification, air removal, and the like.
  • a surfactant is also added to the dispersion medium.
  • surfactants eg Tween 80, Triton X 100
  • Tween 80, Triton X 100 in addition to facilitating the dispersion of the above-mentioned passivating agents, in particular natural oils or/and organic solvent virus inactivating agents, in water, sometimes as an eluent
  • the adsorption of the above passivating agent on the adsorption medium is controlled.
  • the passivating agent is fixed on the adsorbent medium, or substantially by adsorption.
  • the liquid passivating agent can also be directly contacted with the adsorption medium to carry out the adsorption reaction.
  • a powdery adsorption medium for example, activated carbon
  • it is added to the above-prepared passivator dispersion system for adsorption reaction.
  • a bulk adsorption medium for example, an activated carbon filter plate
  • the passivation agent dispersion system prepared above is a mobile phase, and the adsorption medium is used as a stationary phase for a flow adsorption reaction.
  • All binding reactions were carried out using optimized passivator/adsorption media ratios under optimized reaction conditions. These optimizations are performed in accordance with well-known bonding techniques (e.g., adsorption techniques). Conditions for the adsorption reaction include: reactant addition amount, pH, temperature, time, concentration of certain additives (e.g., surfactant, salt, etc.), mobile phase flow rate (at the time of flow adsorption reaction), and the like. Those skilled in the art will recognize that by controlling these conditions, the adsorption reaction is controlled to achieve the desired result (e.g., amount of adsorption). The uniformity of the adsorption reaction is also a priority.
  • adsorption techniques e.g., adsorption techniques.
  • the adsorbent/passivator complex can also be prepared by immobilizing a viral inactivating adsorbent (e.g., C18) on a solid support (e.g., chromatography gel) followed by immobilization of the passivating agent.
  • a viral inactivating adsorbent e.g., C18
  • a solid support e.g., chromatography gel
  • Passivation agents and other substances that are not fixed or weakly adsorbed on the adsorption medium may be washed or prevented by different washing liquids (for example, PBS buffer, preferred concentration of urea solution, alcohol solution, etc.) according to different needs.
  • washing liquids for example, PBS buffer, preferred concentration of urea solution, alcohol solution, etc.
  • the composite prepared above can be used alone as a solid phase medium (for example, A5-A30 in Table 1), or with other components (such as a speed increasing substance, a binder, a solubilizing agent, etc.). After mixing, it is used as a solid phase medium containing an adsorption medium and a passivating agent.
  • a solid phase medium containing an adsorption medium and a passivating agent.
  • A1 adsorption medium/passivator complex and 10% by volume chromatographic gel Sepharose FF, product of Pharmacia
  • A2 contains adsorption medium/passivator complex and 10% by volume Perlite mix.
  • components in the solid phase medium can be passivated according to actual needs according to the undesired reactivity of the medium (for example, forming other solid phases)
  • the component/passivator complex or after mixing with the adsorption medium, is passivated (eg, forming a solid phase media/passivator complex:).
  • Their passivation method is the same as that in the above-described method for preparing an adsorbent medium/passivator compound.
  • A1 is prepared by first forming a solid phase component/passivator complex and then mixing to form a solid phase medium; A2 is first mixed to form a solid phase medium, and then a passivating agent is added to passivate the different components. Prepared.
  • A1 activated carbon powder castor oil A21 AKS5 Tween -80
  • virus inactivating agent adsorption media containing the passivating agent are both an organic solvent virus inactivating agent adsorption medium and a photosensitizer virus inactivating agent adsorption medium (for example, those containing activated carbon and macroporous adsorption resins).
  • the passivating agent content of the solid phase medium (total amount of passivating agent added - unfixed passivating dose) / amount of solid phase medium.
  • the unfixed passivation dose is obtained by well-known correlation measurement techniques.
  • the method for measuring natural fats and oils is a spectrometer method; the method for measuring an organic solvent virus inactivating agent is gas chromatography; the method for measuring a surfactant is gas chromatography; and the method for determining a polypeptide is high pressure liquid chromatography; The method of determination is high pressure liquid chromatography; and the like.
  • the content of the passivating agent is greater than 0.05 ⁇ 1/ ⁇ 3 and individually greater than 0.4 mmol/cm 3 .
  • the specific adsorption amount of the virus inactivating agent (the total amount of the virus inactivating agent added - the unadsorbed virus inactivating dose y the volume of the solid phase medium).
  • the organic solvent virus inactivating agent (TnBP) and the photosensitizer virus inactivating agent (methylene blue) are divided into Do not be used to determine specific adsorption.
  • the measurement of methylene blue uses a well-known spectrophotometer method.
  • the measurement of TnBP uses a well-known gas chromatography.
  • the amount of adsorption of the virus inactivating agent is measured by a known method for measuring the dynamic adsorption amount.
  • the apparatus used for the measurement was a column container (volume 10 ml) or a filter (volume 10 ml) respectively containing the solid phase medium and the control medium prepared in the present example, and 100 ml of the virus inactivating agent solution (methylene blue concentration 100 g/ml) was measured.
  • the TnBP concentration of 10 mg / ml was passed through the apparatus at a linear velocity of 0.3 cm / min.
  • the adsorption amount of the solid phase medium A1-A35 to the photosensitizer virus inactivating agent (methylene blue) is greater than O.Olmmol/cm 3 , and the individual is greater than 0.02 mmol/cm 3 , and sometimes even greater than 0.1 mmol/cm.
  • the adsorption amount of the organic solvent virus inactivating agent (TnBP) of the solid phase media A1-A30 and A36-A39 is greater than 0.05mmol/cm 3 , and the individual is greater than O.lmmol/cm 3 , individual even greater than 0.3mmol / cm 3 ; A1-A30 adsorption of iodine greater than lmmol / cm 3 .
  • some of the above solid phase media especially those containing activated carbon have similar results for the specific adsorption of other photosensitizer virus inactivating agents, such as psoralen.
  • A33-A35 can also be used as a leukocyte-removing medium.
  • the measurement of side effects includes measuring the amount of non-specific adsorption or / and APTT (human plasma fraction prothrombin activity).
  • the amount of non-specific adsorption (indicating the total amount of reagent added - the total amount of indicator reagent not adsorbed) / the volume of the solid phase medium.
  • human albumin (sample C) (Tiantan Biological Products Co., Ltd.) was used as a non-specific adsorption indicating reagent; partial prothrombin activity time of human plasma (sample D) (abbreviated as APTT in the present invention) Used as an indicator of changes in the coagulation system.
  • the sputum kit was purchased from the Chengdu Institute of Blood Transfusion, Chinese Academy of Medical Sciences.
  • the measurement of side effects is also carried out by a known dynamic reaction measurement method.
  • the ratio of human albumin (5%) or human plasma (5.5% protein) to the solid medium was between 3:1 and 5:1.
  • the adsorption medium/passivator composite prepared in this example has a significant decrease in side effects, which is reflected as: A).
  • the albumin adsorption amount decreases by more than 25%, and the individual decreases by more than 50%.
  • the increase in human plasma APTT is more than 30%, and the individual drop is more than 100% ( For example, albumin/adsorption medium complex, vegetable oil/adsorption medium complex, sugar/adsorption medium complex, etc.).
  • the solid phase medium prepared in this example is an organic solvent virus inactivating agent/adsorption medium/passivator compound.
  • a virus inactivating agent solution or suspension is prepared according to a known technique using PBS buffer as a dispersed phase, for example: TnBP/Triton X 100/water solution (5% TnBP, 5% Triton X 100); TnBP/spray Warm 80/water solution (3% TnBP, 10% Tween 80 concentration); ⁇ -propiolactone/Tween 80/water solution (0.5% ⁇ -propiolactone);
  • the virus inactivating agent in the liquid phase is adsorbed on the solid phase adsorption medium by a conventional solid-liquid phase adsorption reaction condition.
  • the fixing method is the same as the method of "fixing the passivating agent to the adsorption medium" in the embodiment 1.1.
  • the preparation method of the virus inactivating agent/adsorption medium complex in this embodiment is the same as the method of "fixing the virus inactivating agent to the adsorption medium" in the present embodiment (1) (e.g., iodine-PVPP filter plate).
  • the method of fixing the passivating agent to the virus inactivating agent/adsorption medium composite is the same as the method of "fixing the passivating agent to the adsorption medium" in the embodiment 1.1.
  • the organic solvent virus inactivating agent/adsorption medium/passivator complex (for example, B1-B3 in Table 2) in this embodiment may be used alone or in combination with other components (for example, a speed increasing substance, a binder, The solubilizer, etc.) is used as a solid phase medium containing the virus inactivating agent after mixing.
  • the passivation method may also be first mixed to form a solid phase medium, followed by passivation to passivate, or passivate the different components and then mix them.
  • Virus inactivation medium prepared in the present embodiment the measured efficiency of inactivated virus into which ⁇ 1 described above, measurement was carried out at room temperature, less than the linear flow rate of lml / cm / min, the volume of biological fluid with a solid support is less than 100.
  • the virus inactivating agent/adsorption medium/passivator complex prepared in this example has an effective virus inactivating function.
  • the virus inactivating medium prepared in this example has the same method of determining the passivating agent content, the virus inactivating agent content, and the solid phase medium side effect as in the corresponding method in the first embodiment.
  • the virus inactivating medium prepared in this example in Table 2 has a passivating agent content of more than 0.05 mol/cm 3 and an individual content of more than 0.4 mmol / cm 3 .
  • the virus inactivation efficiency was not significantly changed by passivation, but the side effects were significantly decreased: A).
  • the albumin adsorption amount (mg/cm 3 ) decreased by more than 15%, and decreased individually. More than 30%; B).
  • the increase in human plasma APTT is more than 20%, and the individual is reduced by 50%. the above.
  • the virus inactivating agent comprises an organic solvent virus inactivating agent
  • the passivating agent comprises an organic solvent virus inactivating agent
  • C The content of the organic solvent virus inactivating agent in the solid phase medium is greater than 0.1 mmol / cm 3 , or even greater than 0.3 mmol / cm 3 (for example, 0.40 mmol / cm 3 ) 0 at this time, the organic solvent virus inactivating agent can It can be used as a virus inactivating agent for effective virus inactivation, and can also be used as a passivating agent for a virus inactivating agent adsorption medium (for example, activated carbon) to minimize side effects.
  • a virus inactivating medium containing passivating agent (2)
  • a solid phase virus inactivating agent/passivator complex was prepared.
  • the solid phase virus inactivating agents used were Cuno-Zetaplus VR virus-killing filter plates and Seitz-iodine-PVPP filter plates.
  • the method of fixing the passivating agent to the solid phase virus inactivating agent is the same as the method of "fixing the passivating agent to the adsorption medium" in Example U.
  • the preparations are listed in Table 2.
  • the virus inactivating medium prepared in this example has the same method for determining the virus inactivation efficiency, the depressant content, and the side effects, respectively, in accordance with the corresponding measurement methods of the examples.
  • the solid phase virus inactivating agent/passivator complex prepared in this example in Table 2 has a passivating agent content of more than 0.05 ⁇ 1/ ⁇ 3 . Compared with the control solid phase virus inactivating agent, the virus inactivation efficiency was not significantly changed by passivation, but the side effects were significantly decreased: ⁇ ).
  • Example 1 4 virus inactivating agent adsorption medium containing deactivating agent / leukocyte solid phase medium
  • the leukocyte-removing medium-virus inactivating agent adsorption medium used is: glass fiber, cotton fiber, polyester fiber, polyurethane nonwoven fabric, Seitz-Supradur 100 polypropylene fiber filter plate
  • the passivating agent used is the above passivating agent (for example, glycine, dextran 40/glucose, and mixtures thereof).
  • the passivating agent/viral inactivating agent adsorption medium-de-whitening cell solid phase medium composite prepared in this embodiment is prepared in the same manner as the adsorption medium/passivator complex in the embodiment 1.1, and the adsorption reaction is in the leukocyte-removing cell.
  • the medium-viral inactivating agent is adsorbed between the adsorption medium and the passivating agent, and the method for determining the content of the passivating agent and the side effect of the solid phase medium is the same as that of the corresponding method in Example 1.1, respectively.
  • the preparation of the present embodiment has a leukocyte removal rate of over 99%, and a methylene blue removal rate of over 95%, and the side effects thereof are significantly lower than the corresponding leukocyte-removing medium without a passivating agent: A).
  • Albumin The amount of adsorption (mg/cm 3 ) decreased by more than 15%; B).
  • the increase in human plasma APTT decreased by more than 20%.
  • the solid phase medium used is selected from the passivated solid phase medium prepared in Example 1, especially the examples 1.1-1.4 therein.
  • the apparatus of this embodiment is obtained by two methods: A) loading a prepared passivated solid phase medium into the chamber; B) loading the solid phase medium into the chamber and then passivating it with a passivating agent,
  • the passivation method is the same as the method of "fixing the passivating agent to the adsorption medium" in Example 1.
  • the apparatus prepared in this embodiment has the same identification method as the corresponding identification method of the passivated solid phase medium in Embodiment 1.
  • its functions e.g., specific adsorption capacity, leukocyte removal rate, virus inactivation efficiency
  • side effects are consistent with the function and side effects of the passivated solid phase medium contained therein, respectively. That is, the function is not significantly reduced by the addition of a passivating agent, and its side effects are significantly reduced.
  • the solid phase medium in the filter is selected from the group consisting of a passivating agent/viral inactivating agent adsorption medium (fiber or/and filter medium).
  • a passivating agent/viral inactivating agent adsorption medium fiber or/and filter medium.
  • passivating agent/polystyrene fiber composite passivating agent/polypropylene fiber composite
  • passivating agent/activated carbon filter plate composite and the like.
  • the solid phase medium in the virus inactivating agent removal column is selected from the group consisting of a passivating agent/viral inactivating agent.
  • Medium (particle) composite For example, passivating agent/polystyrene macroporous adsorption resin composite, passivating agent/activated carbon powder composite, and the like.
  • the solid phase medium in the virus inactivating filter is: 1) a virus inactivating medium immobilized with a passivating agent (for example, a high content of TnBP/earth temperature 80/activated carbon filter plate composite); a virus inactivating medium immobilized with a passivating agent and a virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, a high content of TnBP/earth temperature 80/activated carbon filter plate complex and a sugar-amino acid purifying agent/activated carbon) a filter plate complex; and 3) a virus inactivating medium and a virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, TnBP/earth temperature 80/activated carbon filter plate complex and sugar-amino acid purifying agent/activity Carbon filter plate composite :).
  • a virus inactivating medium immobilized with a passivating agent for example, a high content of TnBP/earth temperature 80/
  • the solid phase medium in the virus inactivation column is: 1) a virus inactivating medium immobilized with a passivating agent (for example, a high content of TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite) 2).
  • a passivating agent for example, a high content of TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite
  • Virus inactivation medium with passivator immobilized and virus inactivating agent adsorption medium with or without passivating agent eg high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin complex
  • passivating agent eg high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin complex
  • sugar-amino acid purifier/activated carbon filter plate composite or high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite and styrene-containing polymer adsorption resin :
  • virus Inactivation medium and virus inactivating agent adsorption medium to which a passivating agent is immobilized for example, styrene-containing polymer adsorption resin/TnBP/earth temperature 80 composite and passivating agent/styrene-containing polymer adsorption resin composite.
  • the solid phase medium in the leukocyte-removing filter is selected from the group consisting of a passivating agent / a leukocyte-removing medium / a virus inactivating agent adsorption medium complex.
  • a filter cartridge is packed with a T 2600 filter plate, 4 layers of passivator/Seitz-Supradur 100 filter plate composite, a 0.4 ⁇ filter at the outlet end, and then pumped into passivation with 8% dextran. Hours, dry and disinfected for use.
  • This filter can be used to remove leukocytes and viral inactivating agents such as methylene blue.
  • the solid phase medium comprises the above-mentioned photosensitizer physical adsorption fiber, such as natural fiber, polyamide fiber, polyurethane fiber and filter plate (for example, Supmdur 100, Supradur 500 Eco l000, Permadur 0/400A T2600 described above). , Bio20, Bio40, Bio60, etc.).
  • the apparatus prepared in this embodiment has the same identification method as the corresponding apparatus prepared in the second embodiment.
  • the apparatus prepared in this example has a similar photosensitizer virus inactivating agent removal ability and has significantly lower side effects than the existing apparatus (for example, a device containing activated carbon or an inorganic adsorption medium).
  • the existing apparatus for example, a device containing activated carbon or an inorganic adsorption medium.
  • protein adsorption is 15% lower or / and APTT value is lower than 15%
  • Lower particle contamination for example, the diameter of the biological fluid flowing through the treatment system visible under the microscope is greater than
  • has a lower particle count of 15% or more).
  • Example 3.1 Dye or / and psoralen photosensitizer virus inactivating agent removal filter
  • the apparatus prepared in this example (including 3.1.1 and 3 ⁇ 2) is suitable for the removal of dyes or/and psoralen-type photosensitizer virus inactivating agents.
  • the photosensitizer virus inactivating agents used are methylene blue, phthalocyanine dye, hypericin, cyanine dye and psoralen.
  • Example 3.1.1 Virus inactivating agent removal filter containing photosensitizer virus inactivating agent adsorption fiber
  • the virus inactivating agent adsorbing fiber is a fiber of an organic polymer physical adsorbent containing a dye or/and a psoralen photosensitizer virus inactivating agent, such as ultrafine polypropylene oil absorbing fiber or/and poly Polyurethane fiber.
  • the fiber loading thickness is 5 cm.
  • Embodiment 3.1.2 Virus Inactivating Agent Removal Filter Containing Photosensitizer Virus Inactivating Agent Adsorbing Fiber and Adsorbing Particles
  • the adsorbing particles are dye-containing or/and psoralen-based photosensitizer virus inactivating agents.
  • Granules such as polypropylene matrix resins (e.g., HP2MG series), styrene containing macroporous adsorption resins (e.g., Amberlite XAD-7HP Amberlite XAD-16), and the like.
  • the fibers used are contained in the filter material used, such as various Seitz products (KS80, K900, Supradur 80 > P30, Eco 1000, Permadur 0/400A, T2600, Bio20, Bio40, Bio60, Supradur 100, and many more).
  • These filter plates have an adsorption effect on photosensitizer virus inactivating agents, particularly dye-based photosensitizers.
  • the filter has a diameter of 1.5 cm, the macroporous adsorption particles are packed to a thickness of 3 cm, and a 3.5 mm thick filter plate is loaded under the adsorbed particles.
  • these filter plates prevent the macroporous adsorption particles from falling off and contaminating the biological fluid to cause side effects.
  • a device prepared in accordance with this embodiment (including 3.2.1-3.2.4) is suitable for removing a dye-based photosensitizer virus inactivating agent such as methylene blue.
  • Example 3.2.1 Virus inactivating agent removal filter containing polyolefin fiber filter plate
  • the virus inactivating agent adsorption medium is selected from the group consisting of a polyolefin fiber filter material of an organic polymer physical adsorbent containing a dye-based photosensitizer virus inactivating agent, such as the above-mentioned Eco 1000, Permadur 0/400A, T2600, Supradur. 80> Supradur 100, Supradur 500.
  • the filter plate is filled with more than 3 layers (each thickness is 3.5mm).
  • the virus inactivating agent adsorption medium is selected from the group consisting of natural fiber-containing filter materials, such as various Seitz company products (e.g., Bio20, Bio40, Bio60, etc.).
  • the filter plate is filled with more than 3 layers (each thickness is 3.5mm).
  • Example 3.2.3 Virus inactivating agent removal filter containing natural fiber
  • the virus inactivating agent adsorption medium is selected from natural fibers such as cotton wool fibers, silk fibers or wood fibers.
  • the virus inactivating agent adsorption medium is selected from the group consisting of synthetic fibers containing organic polymer physical adsorbents, such as polyurethane fibers or polystyrene fibers.
  • Example 3.3 Dye-like virus inactivating agent removal / removal of leukocyte filter
  • Some of the devices prepared as described above in Example 3 can also be used for dye virus inactivating agents to remove and remove leukocytes.
  • cotton fibers, polyesters, polyurethanes, and the like are both a dye-based photosensitizer virus inactivating agent adsorption medium and a leukocyte-removing material.
  • the apparatus prepared in this embodiment wherein the solid phase medium contains the above organic solvent physical adsorption medium.
  • the apparatus prepared in this example was identified in the same manner as the apparatus prepared in Example 2.
  • the devices prepared in this example have similar virus inactivating agent removal/and virus inactivation efficiencies compared to existing devices, with significantly lower side effects (eg, with activated carbon or inorganic adsorption). Comparison of medium devices): Protein adsorption is 15% lower or / and APTT value is lower than 15%.
  • the apparatus of this embodiment is prepared by charging the above organic solvent physical adsorption medium into a filter drum or a hollow column.
  • Organic solvent physical adsorption medium used including fibers (for example, ultrafine polypropylene oil absorbing fibers), filter plates (for example) or/and macroporous adsorption resins (such as Supradur 80), polyphenylene-containing porous particles (Amberlite XAD-7HP) AmberliteXAD-16, etc.).
  • the apparatus of the present embodiment is prepared by charging an organic solvent virus inactivating medium into a filter drum or a hollow column.
  • the organic solvent virus inactivating medium contains the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium.
  • the preparation method used in this method is the same as the method of "fixing the virus inactivating agent to the adsorption medium" in Example 1.2, wherein: the adsorption medium used is selected from the above organic solvent physical adsorption medium (for example, used in Example 4.1).
  • the organic solvent physical adsorption medium the virus inactivating agent used is S (for example, TnBP or ⁇ -propiolactone) or SD (for example, ⁇ /earth temperature-80, TnBP/Triton X100, or TnBP/alcohol).
  • S for example, TnBP or ⁇ -propiolactone
  • SD for example, ⁇ /earth temperature-80, TnBP/Triton X100, or TnBP/alcohol.
  • Another preparation method is: loading the above organic solvent physical adsorption medium into a filter drum or a hollow column, pre-washing, and then using an organic solvent dispersion system to flow through the adsorption medium at an optimized speed (for example, 0.1-lcm/min) to remove the organic solvent.
  • the virus inactivating agent is fixed and then the unfixed material is washed out.
  • the preparation method of the organic solvent dispersion system is the same as the method of "preparation of the organic solvent dispersion system" in Example 1.1.
  • Organic solvent Examples of the dispersion system include the following solutions in appropriate concentration ratios: TnBP/earth temperature-80 solution, TnBP/Triton 5Q00 solution, TnBP/alcohol solution, and the like.
  • the content of the organic solvent-inactivated medium containing the organic solvent inactivating agent may vary depending on the needs. For example, if the volume of the biological fluid to be treated is 250 ml, the TnBP content of some devices is greater than 7.5 mg, and the TnBP content of some devices is equal to or less than 7.5 mg, or even equal to or less than 2.5 mg. It corresponds to the content of the organic solvent virus inactivating agent respectively greater than 3 X (lOppmX volume of the single-part blood component ml), equal to or less than 3 X (lOppmX volume of the single-part blood component ml), And equal to or less than (lOppmX the volume of the single-part blood component ml).
  • the apparatus of this embodiment is prepared by separately loading an organic solvent virus inactivating medium and an organic solvent virus inactivating agent adsorption medium into a filter drum or a hollow column.
  • the organic solvent virus inactivation medium is placed near the inlet of the chamber, and the organic solvent virus inactivating agent adsorption medium is placed near the outlet;
  • the organic solvent virus inactivating agent is adsorbed The medium is placed near the inlet of the chamber and near the outlet, and the organic solvent virus inactivating medium is placed therebetween.
  • the organic solvent virus inactivating device may be connected in series with the organic solvent virus inactivating agent removing device.
  • the solid phase medium used is a combination of one of the following: 1) an organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (refer to the implementation) Example 4.2), and the above organic solvent physical adsorption medium (refer to Example 4.1); 2).
  • Other organic solvent virus inactivating medium for example, the organic solvent virus inactivating medium prepared in Example 1
  • the above organic solvent physical adsorption medium Reference Example 4.1
  • Organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and organic solvent virus inactivating agent immobilized on the adsorption medium (refer to Example 4.2), and other organic solvent viruses
  • the active agent adsorption medium for example, the organic solvent virus inactivating agent adsorption medium prepared in Example 1).
  • the apparatus of the present embodiment is an organic solvent virus inactivating medium, an organic solvent virus inactivating agent adsorption medium, and a photosensitizer virus inactivating agent adsorption medium (or an organic solvent virus inactivating medium and an organic solvent virus inactivating agent/photosensitive).
  • the agent virus inactivating agent adsorption medium is prepared by separately loading into a filter drum or a hollow column. There are two methods of filling: A). The organic solvent virus inactivating medium is placed near the inlet of the chamber; B) The organic solvent virus inactivating agent medium is placed near the middle of the chamber. Alternatively, it may be formed by an organic solvent virus inactivating device, an organic solvent virus inactivating agent removing device, and a photosensitizer virus inactivating agent removing device.
  • the solid phase medium used is a combination of one of the following: 1) an organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (see Test Example 4.2), the above organic solvent physical adsorption medium (refer to Example 4.1), and the above photosensitizer physical adsorption fiber (Reference Example 3); 2).
  • Organic solvent virus inactivation medium for example, prepared in Example 1
  • Organic solvent virus inactivating medium for example, prepared in Example 4.1
  • organic solvent physical adsorption medium for example, 4.1
  • photosensitizer physical adsorption fiber for example, the above photosensitizer prepared in Example 1
  • photosensitizer adsorption medium for example, the photosensitizer prepared in Example 1
  • Virus inactivating agent adsorption medium 3
  • Organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and organic solvent virus inactivating agent immobilized on the adsorption medium (refer to Example 4.2), and other organic solvent viruses Inactivator adsorption medium (for example, organic solvent virus inactivating agent adsorption medium prepared in Example 1), and photosensitizer adsorption medium (for example, the above photosensitizer physical adsorption fiber or the photosensitizer virus inactivating agent adsorption medium prepared in Example 1) 4).
  • An organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (Reference Example 4.2) And a photosensitizer / organic solvent virus inactivating agent adsorption medium (e.g., a polymer containing styrene macroporous adsorption resin, passivating agent prepared in Example 1 / activated carbon composite, etc.).
  • a photosensitizer / organic solvent virus inactivating agent adsorption medium e.g., a polymer containing styrene macroporous adsorption resin, passivating agent prepared in Example 1 / activated carbon composite, etc.
  • Filter F6 A15 (Table 1) A28 (Table 1) In addition to SD agent / photosensitizer virus inactivating agent and its derivatives Filter F7 A20 (Table 1) A28 (Table 1) In addition to SD agent / photosensitizer virus inactivating agent and its derivatives
  • Filter F9 A9 (Table 1) Seitz- Bio 40, in addition to SD agent / photosensitizer virus inactivating agent and its polyester oil-absorbing fiber derivative / leukocyte
  • the function and side effects of the filter in this embodiment are consistent with the functions and side effects of the solid phase medium contained therein.
  • Example 6 Example of a single blood component kit of the present invention
  • the term "single-part blood component processing device” refers to a biological liquid processing device in which a single-part blood component is a biological liquid.
  • the single-part blood component processing device of the present invention includes single-part plasma Virus inactivation device, single-part platelet virus inactivation device (for example, using psoralen as a virus inactivating agent), etc. Based on the above device, several other devices may also be derived. For example, by interacting with other processing systems (eg, The blood collection and separation processing system are connected to form a more integrated device.
  • the kit prepared in this embodiment (including 6.1 to 6.3) contains at least a single blood component treatment device, and is a device prepared in the above embodiment.
  • the kit also contains a viral inactivating agent addition structure, including a liquid phase virus inactivating agent addition structure or a solid phase virus inactivating agent addition structure.
  • a viral inactivating agent addition structure including a liquid phase virus inactivating agent addition structure or a solid phase virus inactivating agent addition structure.
  • liquid phase viral inactivating agents eg, photosensitizer virus inactivating agent solutions
  • deblockable closed structures eg, switches or Fragile medical polystyrene plastic sheet.
  • the kit also contains a viral inactivation liquid reaction site, such as a clear plastic bag for the photosensitizer virus inactivation, and the like.
  • the kit also contains other structures (eg, in addition to leukocyte filters, lotions, lotion addition devices, etc.), where a lotion (eg, saline) is used to place the device, particularly a column device or / The blood components retained on the solid phase medium in the filter device are washed out to reduce loss of blood components.
  • a lotion eg, saline
  • the function and side effects of the kit in this embodiment are consistent with the functions and side effects of the device contained therein (refer to the related embodiments above). If other parts of the kit have unwanted reactivity, follow the instructions. It is necessary to pass a substance selected from the above passivating agent for passivation.
  • the passivation methods include: 1) passivating and reassembling the parts that need to be passivated into devices; 2) passivating the assembled device; 3) passivating and reassembling the parts that need to be passivated The device is then re-passivated for the assembled device.
  • Example 6.1.1 Single Human Blood Component Virus Inactivating Kit Containing Virus Inactivating Agent Adsorption Treatment System
  • the kit prepared in this embodiment comprises a virus inactivating agent adsorption treatment system, i.e., a filter or column containing the virus inactivating agent adsorption medium prepared in the above embodiment.
  • the kit also contains a viral inactivating agent addition device and a virus inactivation reaction site.
  • the working principle is as follows: the single blood component is contacted with the virus inactivating agent added by the virus inactivating agent, and the virus inactivation reaction is carried out at the virus inactivation reaction site, and the biological fluid flows at a preferred flow rate after the reaction is completed.
  • the virus inactivating agent adsorbs the filter or column of the medium and removes the virus inactivating agent or/and its derivative, and if necessary, washes the retained blood component by adding the washing liquid to the structure.
  • Example 6.1.2 Single Human Blood Component Virus Inactivating Kit Containing Virus Inactivation Treatment System
  • the kit prepared in this embodiment comprises a virus inactivation treatment system, i.e., a virus inactivating medium-containing filter or column prepared in the above embodiment.
  • a virus inactivation treatment system i.e., a virus inactivating medium-containing filter or column prepared in the above embodiment.
  • the working principle is as follows: a single human blood component flows through a filter or column containing a virus inactivating medium at a preferred flow rate for virus inactivation (for example, immobilized SD agent treatment, immobilized iodine treatment, or immobilized ion treatment) If necessary, the blood components retained in the treatment system can be washed out by adding the washing liquid to the structure and adding the washing liquid.
  • Example 6.1.3 Single-part blood component virus inactivating kit containing virus inactivation/viral inactivating agent adsorption treatment system
  • the kit prepared in this embodiment comprises a virus inactivation treatment system, that is, a filter or column containing the virus inactivating medium and the virus inactivating agent adsorption medium prepared in the above embodiment.
  • the working principle is as follows: The single blood component is subjected to virus inactivation by a preferred flow rate filter or a virus inactivating medium in the column (for example, immobilized SD treatment, immobilized iodine treatment, or immobilized ion treatment: And flowing through the virus inactivating agent adsorption medium to ensure that the blood component of the effluent filter or column is substantially free of viral inactivating agents. If necessary, the blood components retained in the treatment system can be washed out by adding the washing liquid to the structure.
  • Example 6.1.4 Single Human Blood Component Virus Inactivating Kit Containing Dual Virus Inactivation Treatment System
  • the kit prepared in the present embodiment which comprises the apparatus prepared in Example 5, further comprises a virus inactivating agent addition device and a first virus inactivation reaction site.
  • the working principle is as follows: the single blood component is contacted with the virus inactivating agent added by the virus inactivating agent, and the virus inactivation reaction is carried out at the virus inactivation reaction site, and the biological liquid is flowed through at a preferred flow rate after the reaction is completed.
  • the filter or column prepared in Example 5 was subjected to virus inactivation and virus inactivating agent removal, and the blood components exiting the filter or column were substantially free of viral inactivating agents. If necessary, the blood components retained in the treatment system can also be washed out by adding the washing liquid to the structure to add the washing liquid.
  • Example 6.2 Single Human Blood Component Virus Inactivation/De-Leukocyte Kit comprising a leukocyte-removing and virus inactivating agent adsorption treatment system, that is, the de-leukocyte-containing and virus inactivating agent adsorption medium prepared by the above embodiment (or containing de-white blood cells and virus inactivating medium, or A filter or column that removes white blood cells and virus inactivating media and virus inactivating media.
  • a leukocyte-removing and virus inactivating agent adsorption treatment system that is, the de-leukocyte-containing and virus inactivating agent adsorption medium prepared by the above embodiment (or containing de-white blood cells and virus inactivating medium, or A filter or column that removes white blood cells and virus inactivating media and virus inactivating media.
  • the method of the present embodiment comprising at least: A) providing a biological fluid; B) providing a processing system, the processing system comprising at least a solid phase medium and a chamber containing the solid phase medium, and At least a portion of the solid phase medium comprises a passivating adsorption medium; C). flowing the biological fluid through the processing system for viral inactivation or viral inactivating agent adsorption.
  • the treatment system used in this example is selected from the corresponding apparatus comprising the passivated adsorption medium prepared in Example 2, 5 or 6.
  • Example 7.1.1 Example of inactivation of virus by single blood component
  • the biological fluid used is a single-part platelet;
  • the virus inactivating agent used is psoralen (4,-aminomethyl-4,5,8-trimethyl psoralen);
  • a passivating agent for example, polyester oil absorbing fiber, glass fiber and super Fine polypropylene oil absorbing fiber.
  • the virus inactivation method is: adding the virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing virus inactivation at the virus inactivation reaction site (appropriate concentration of psoralen, room temperature, illumination 30) Minutes), then the biological fluid is introduced into the device and the virus inactivating agent is removed by contact with the virus inactivating agent adsorption medium therein.
  • the method of this example is also effective in removing viral inactivating agents (e.g., more than 90% of psoralen removal) with a smaller side effect, as compared to a method using a similar device containing a conventional adsorption medium (e.g., activated carbon).
  • a conventional adsorption medium e.g., activated carbon
  • the protein loss was 10-20%
  • the platelet loss was 7%
  • the platelet morphology score was decreased by 16%.
  • the protein loss was only 5%.
  • the platelet loss was 3%, and the platelet morphology score did not decrease.
  • the biological fluids used are: 1. multi-part combined plasma, 2. human plasma fraction containing human fibrinogen (for example, component I or cryoprecipitate in cold alcohol precipitation), 3. Human plasma separation component (eg, cryoprecipitate) containing human coagulation factor VIII, 4, human plasma separation component containing human coagulation factor IX (eg, DEAE-Sephadex washout component of cryoprecipitate supernatant), 5. Human plasma separation component of human prothrombin complex (PCC) (eg DEAE-Sephadex washout component of cryoprecipitate supernatant), 6. Human plasma fraction containing gamma globulin (eg cold alcohol precipitation method) Component II precipitation:), 7. Recombinant Alpha interferon (interferon concentration 1.15 million IU/ml, pH 7.0), 8. Positive human serum reference.
  • human plasma separation component eg, cryoprecipitate
  • human coagulation factor VIII for example, component I or cryoprecipitate in cold alcohol precipitation
  • the virus inactivating agent used is an S/D agent (for example, TnBP/Triton X100, TnBP/spit Temperature 80, or diethyl ether / Triton X100);
  • the apparatus used is selected from the adsorption filter prepared in Example 2.1 (the size of which varies depending on the amount of biological liquid), and the adsorption filter contains an organic solvent virus with a passivating agent immobilized thereon.
  • Living agent adsorption medium for example, styrene-containing polymer macroporous adsorption resin or activated carbon filter plate).
  • the virus inactivation method in this embodiment is: adding the virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing virus inactivation at the virus inactivation reaction site (0.3-1% S, or 0.3-1%) S and 0.3-1% D; 30 ° C; 4 hours), then the biological liquid is introduced into the above device, and the organic solvent virus inactivating agent is removed by contact with the virus inactivating agent adsorption medium therein.
  • the method of this example is also effective in removing viral inactivating agents (e.g., more than 90% of TnBP removal) compared to methods using similar devices containing conventional adsorption media (e.g., activated carbon), with side effects being much smaller.
  • viral inactivating agents e.g., more than 90% of TnBP removal
  • conventional adsorption media e.g., activated carbon
  • the virus inactivation method in this example results in an increase in plasma APTT value of less than 30% compared to existing methods using activated carbon, such that the biological activity of other biological fluids (eg, fibrinogen, coagulation factor VIII)
  • the loss rate of coagulation factor IX and total protein is 30% or less.
  • the biological fluid used is a single-part plasma; the apparatus used is selected from the kit prepared in Example 6, which comprises the virus inactivating apparatus prepared in Example 2.2, which respectively contains the organic solvent virus inactivating agent/adsorption medium. / Passivator complex, Crnio-Zetaplus VR virus filter plate and Seitz-iodine-PVPP filter plate.
  • the virus inactivation treatment in the present embodiment can enable the plasma product factory to perform virus inactivation after the pulping to reduce the risk of virus contamination in the blood storage, transportation and production process, and improve the product. Virus security.
  • the virus inactivation method in this embodiment is: a biological fluid at room temperature is passed through the above apparatus at a preferred flow rate (e.g., a linear velocity of 0.1 - 1.5 cm / min:) to carry out virus inactivation. If necessary, wash the device including the solid phase media with sputum.
  • a preferred flow rate e.g., a linear velocity of 0.1 - 1.5 cm / min:
  • the method of the present embodiment can equally effectively inactivate the virus (for example, the pattern virus is inactivated by more than 4 logs), and has less unnecessary Reactivity (for example, the APTT value is increased by more than 20%, and the loss rate of fibrinogen, coagulation factor VIII, coagulation factor IX, and total protein is 15% or less).
  • a common adsorption medium for example, activated carbon
  • the biological fluids used are: 1. multi-part combined plasma, 2. human plasma fraction containing human fibrinogen (for example, component I or cryoprecipitate in cold alcohol precipitation), 3 Human plasma separation component of human prothrombin complex (PCC) (eg, DEAE-Sephadex washout component of cryoprecipitate supernatant).
  • the apparatus used is selected from the virus inactivation apparatus prepared in Example 2.2 (the volume of the chamber is different from the volume of the biological liquid), and the adsorbent medium/organic solvent virus inactivating agent/passivator complex is respectively contained. , Cuno-Zetaplus VR virus-killing filter plate and Seitz-iodine-PVPP filter plate.
  • the virus inactivation method in this example is the same as in the method of Example 7.2.1, and the comparison with the method using a similar apparatus containing a usual adsorption medium (e.g., activated carbon) is also in agreement with the comparison in Example 7.2.1.
  • a usual adsorption medium e.g., activated carbon
  • the biological fluid used is a single-part plasma;
  • the virus inactivating agent used is selected from the photosensitizer virus inactivating agent (for example, 4,-aminomethyl-4,5,8-trimethyl psoralen)
  • the kit used was selected from the kit prepared in Example 7, which contained the leukocyte/virus inactivating agent removal filter prepared in Example 2.4.
  • the leukocyte removal/viral inactivating agent removal method in this embodiment is: adding a virus inactivating agent to a biological liquid through a virus inactivating agent addition device, and performing virus inactivation at a virus inactivation reaction site (for example, an appropriate concentration of virus) The inactivating agent, room temperature, light for 30 minutes), then the biological liquid is introduced into the above device, and the virus inactivating agent and its derivative are removed by contact with the solid phase medium therein.
  • the method of this example has an equivalent leukocyte removal efficiency and a significant reduction in side effects as compared to a method using a similar device containing a conventional adsorption medium such as glass fiber.
  • Example 8. Virus inactivation method of the present invention (2)
  • the method of the present embodiment comprising at least: A) providing a biological fluid; B) contacting the biological fluid with a viral inactivating agent and performing virus inactivation (eg, an appropriate concentration of virus is eliminated) Living agent, room temperature, illumination for 30 minutes), wherein the virus inactivating agent comprises a photosensitizer virus inactivating agent; C) providing a treatment system, the treatment system containing at least a virus inactivating agent adsorption medium and containing the solid phase a chamber of the medium, wherein the virus inactivating agent adsorption medium contains a photosensitizer physical adsorption fiber; D). flowing the biological liquid treated by the step B) through the treatment system to remove the photosensitizer virus inactivating agent adsorption and its derivative .
  • virus inactivation eg, an appropriate concentration of virus is eliminated
  • Living agent room temperature, illumination for 30 minutes
  • the virus inactivating agent comprises a photosensitizer virus inactivating agent
  • C) providing a treatment system the treatment system containing
  • the biological fluid used was a single-part plasma; the treatment system used was the apparatus prepared in Example 7, which contained a device (filter or column) selected from the preparation of Example 3.
  • the method of the present embodiment can effectively remove inactivated virus agents (e.g., more than 95%) with less unwanted reactivity (e.g., compared to methods using similar devices containing conventional adsorption media (e.g., activated carbon).
  • the APTT value is increased by more than 20%, and the loss rate of fibrinogen, coagulation factor VIII, coagulation factor IX, and total protein is 15% or less).
  • the virus inactivating agent used is selected from the group consisting of dyes or psoralen photosensitizer virus inactivating agents (for example, methylene blue or 4,-aminomethyl-4,5,8-trimethyl) Psoralen:)
  • the device used was selected from the kit prepared in Example 6, which contained the adsorption filter prepared in Example 3.1.
  • the virus inactivating agent used is selected from the group consisting of a dye-based photosensitizer virus inactivating agent (for example, methylene blue);
  • the apparatus used was selected from the kit prepared in Example 6, which contained the adsorption filter prepared in Example 3.2.
  • the virus inactivating agent used is selected from a dye-based photosensitizer virus inactivating agent (e.g., methylene blue); and the apparatus used is selected from the kit prepared in Example 6, which contains the adsorption filter prepared in Example 3.3.
  • a dye-based photosensitizer virus inactivating agent e.g., methylene blue
  • the apparatus used is selected from the kit prepared in Example 6, which contains the adsorption filter prepared in Example 3.3.
  • the virus inactivation method in this embodiment includes at least: A) providing a biological fluid; B) providing a treatment system, the treatment system comprising at least a solid phase medium and a chamber containing the solid phase medium Wherein the solid phase medium comprises a virus inactivating medium or/and a virus inactivating agent adsorption medium, and: the virus inactivating medium comprises at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon, The virus inactivating agent adsorption medium comprises at least an organic solvent physical adsorption medium; C). flowing the biological liquid through the treatment system for virus inactivation or/and virus inactivating agent adsorption.
  • the treatment system used in this example was selected from the apparatus prepared in Example 4.
  • Example 9.1.1 Example of inactivation of virus by single blood component
  • the biological fluid used was a single-part plasma; the apparatus used was selected from the kit prepared in Example 6, which contained the apparatus prepared in Example 4.1.
  • the virus inactivation method in this example is the same as the virus inactivation method in Example 7.1.1, and the virus inactivating agent removal efficiency and side effect reduction are also the same.
  • Example 9.1.2 Examples of virus inactivation of other biological fluids
  • the biological fluid used is the same as that used in the embodiment 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.1 (the volume of the chamber varies depending on the volume of the biological fluid).
  • the virus inactivation method in this embodiment is the same as the virus inactivation method in Example 7.1.2, and the virus inactivating agent removal efficiency and side effect reduction are also the same.
  • Example 9.2.1 Example of inactivation of virus by single blood component
  • the biological fluid used was a single-part plasma; the apparatus used was selected from the kit prepared in Example 6, which contained the apparatus prepared in Example 4.2.
  • the virus inactivation method in this example is the same as the virus inactivation method in Example 7.2.1, and the virus inactivation efficiency and side effect reduction are also the same.
  • Example 9.2.2 Examples of virus inactivation of other biological fluids
  • the biological fluid used is the same as the biological fluid used in Example 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.2 (the volume of the chamber is different from the volume of the biological fluid).
  • the virus inactivation method in this embodiment is the same as the virus inactivation method in Example 7.2.2, and the virus inactivation efficiency and side effect reduction are also the same.
  • Example 9.3 Method for Removal of Immobilized Organic Solvent Virus Inactivation/Organic Solvent Virus Inactivating Agent Example 9.3.1 Single Human Blood Component Virus Inactivation Example
  • the biological fluid used is a single-part plasma; the apparatus used is selected from the kit prepared in Example 6, which contains the filter or column prepared in Example 4.3.
  • the virus inactivation method in this embodiment is: flowing the biological fluid at room temperature through the above device at a preferred flow rate, performing virus inactivation and removing the organic solvent virus inactivating agent which may fall off.
  • the virus inactivation efficiency and side effects in the method of this example were the same as in the method of Example 9.2.2, and the content of the organic solvent in the biological liquid collected from the outlet of the apparatus used was less than 10 ppm.
  • the biological fluid used is the same as that used in the embodiment 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.3 (the volume of the chamber varies depending on the volume of the biological fluid).
  • the virus inactivation method in this embodiment is: flowing the biological fluid at room temperature through the above device at a preferred flow rate, performing virus inactivation and removing the organic solvent virus inactivating agent which may fall off.
  • the virus inactivation efficiency, side effects, and organic solvent content of the collected biological fluids in the method of this example were consistent with the method of Example 9.3.1.
  • the method of virus inactivation in this embodiment comprises at least: A) providing a biological fluid; B) performing a first viral inactivation of the biological fluid; C) providing a treatment system, The treatment system comprises at least a virus inactivating medium and a chamber containing the solid phase medium, and wherein the virus inactivating medium contains at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon; D).
  • the biological fluid inactivated by the first virus flows through the treatment system for a second viral inactivation.
  • the biological fluid used is a single blood paddle.
  • the apparatus used is selected from the kit prepared in Example 6, which comprises a device selected from the preparation of Example 4.4 or Example 5.
  • the photosensitizer virus inactivating agent used is selected from the group consisting of dyes or psoralen photosensitizer virus inactivating agents (for example, methylene blue or 4'-aminomethyl-4, 5, 8-3 Methyl psoralen).
  • the treatment system used is selected from the apparatus prepared in Example 4.4 or Example 5, in particular, a photosensitizer virus inactivating agent adsorption medium, an organic solvent virus inactivating medium, and an organic solvent virus inactivating agent adsorption medium.
  • the virus inactivating medium comprises at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon.
  • the virus inactivation method is: adding the photosensitizer virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing the first virus inactivation at the virus inactivation reaction site (for example, an appropriate concentration) Virus inactivating agent, room temperature, light for 30 minutes), then let the biological fluid enter the above device, remove the photosensitizer virus inactivating agent by contact with the solid phase medium therein and carry out the second virus inactivation, and remove the photosensitizer virus Inactivating agent and organic solvent virus inactivating agent.
  • the virus inactivation reaction site for example, an appropriate concentration
  • Virus inactivating agent room temperature, light for 30 minutes
  • the side effects are controlled at an acceptable level due to the use of a solid phase medium having a lower side effect.
  • the APTT value is kept within 47 seconds, and the loss rates of fibrinogen, coagulation factor VIII, and coagulation factor IX are also less than 30%, respectively.
  • the apparatus used is selected from the kit prepared in Example 6, which comprises the filter or column prepared in Example 2.2 or 4.3, which contains an organic solvent virus inactivating medium and an organic solvent virus inactivating agent adsorption medium.
  • the method for inactivating the virus in this embodiment is: adding a known plasma active protective agent (for example, sugar, sodium valerate, calcium, etc.) to the biological liquid through the protective agent adding device, and performing virus at the virus inactivation reaction site.
  • a known plasma active protective agent for example, sugar, sodium valerate, calcium, etc.
  • Inactivate for example, a suitable concentration of protective agent, 52 ° C, 3 hours
  • the biological liquid enter the above device, inactivate the organic solvent virus by contact with the solid phase medium and remove the organic solvent that may fall off Active agent.
  • the side effect of at least the second virus inactivation is small due to the use of a solid phase medium having a lower side effect.
  • the virus inactivation method in this embodiment at least comprises: A) providing a biological fluid; B) providing the device prepared in Example 2, comprising the virus inactivating medium or/and a virus inactivating agent adsorption medium; The biological fluid is passed through the treatment system for virus inactivation or/and viral inactivating agent adsorption, and leukocyte removal.

Abstract

A method of inactivating virus, at least comprising: A) providing bio-fluid; B) providing the treatment system; and C) contacting the bio-fluid with the treatment system, wherein the treatment system at least contains solid-phase medium and the chambers containing the solid-phase medium, which at least comprises: a) the adsorptive medium of the viral inactivator; and /or b) the viral inactivating medium, which comprises the viral inactivator and the adsorptive medium of the viral inactivator, which comprises: (a) deactivation adsorptive medium, which at least contains the adsorptive medium and the deactivator fixed on the surface of the medium; and/or (b) physical adsorptive medium, which comprises photosensitizer physically adsorptive medium as photosensitizer viral inactivator’s adsorptive medium, and organic solvent physically adsorptive medium as organic solvent viral inactivator’s adsorptive medium.

Description

一种病毒灭活方法、 其中所用的处理系统以及装置 技术领域  Virus inactivation method, processing system and device used therein
本发明一般地涉及对生物液体进行处理、 更具体地涉及病毒灭活的方法、 处理系统以及装置。  The present invention relates generally to methods of treating biological fluids, and more particularly to virus inactivation, treatment systems, and devices.
背景技术 Background technique
自二十世纪七十年代以来, 由于生物物质中的病原体 (pathogen)的污染、 例 如病毒 (vims)的污染, 而导致严重说的医疗健康后果的事件, 已成为公众日益关心 的问题。 为此, 科学技术界开发了若干灭活或 /除去这些病原体的技术。 下面以 病毒灭活、 特别是含固-液相反应的病毒灭书活为例, 说明现有技术尚待解决的问 题。  Since the 1970s, incidents of serious medical health consequences due to contamination of pathogens in biological materials, such as viruses (vims), have become a growing concern of the public. To this end, the scientific and technological community has developed several techniques for inactivating or/or removing these pathogens. In the following, the virus inactivation, especially the virus-killing activity containing the solid-liquid phase reaction, is taken as an example to illustrate the problems to be solved in the prior art.
现有的含固-液相反应的病毒灭活技术, 主要基于有机溶剂病毒灭活 (Solvant-Detergent处理, 本发明中简称 SD处理)。 有机溶剂病毒灭活介质含有 机溶剂 S (可另含有去污剂 D)和病毒灭活剂吸附剂, 例如活性碳 (参考中国专利 01123658.2)。 其它含固-液相反应的病毒灭活技术, 还包括固定化烷基磷化合物 处理 (用无机吸附剂作吸附剂, 参考德国专利 DE40(H099A1)、 固定化碘处理 (用 含硅燥土 PVPP滤板作吸附剂, 参考德国 Shenk公司产品介绍)、 等等。 现有技 术的问题是, 所用活性碳、 无机吸附剂或含硅燥土 PVPP滤板具有副作用 (例如 较强的蛋白吸附能力、 凝血因子失活、 血浆 APTT值大幅提高、 等等)。  The existing virus inactivation technique containing solid-liquid phase reaction is mainly based on organic solvent virus inactivation (Solvant-Detergent treatment, referred to as SD treatment in the present invention). The organic solvent virus inactivating medium contains an organic solvent S (which may additionally contain detergent D) and a virus inactivating agent adsorbent such as activated carbon (refer to Chinese Patent 01123658.2). Other virus inactivation techniques containing solid-liquid phase reactions, including immobilization of alkylphosphorus compounds (using inorganic adsorbents as adsorbents, reference to German patent DE40 (H099A1), immobilized iodine treatment (with silicon-containing dry soil PVPP) The filter plate is used as an adsorbent, refer to the product introduction of the German Shenk company, etc. The problem of the prior art is that the activated carbon, the inorganic adsorbent or the silicon-containing dry soil PVPP filter plate has side effects (for example, strong protein adsorption capacity, Inactivation of coagulation factors, a significant increase in plasma APTT values, etc.).
现有用固相介质吸附去除不需要的病毒灭活剂的技术中, 有机溶剂病毒灭 活剂吸附介质分别为无机吸附剂 (例如含珍珠岩的除脂滤板:)、 亲和吸附剂 (例如 C-18 亲油树脂)、 活性碳。 现有技术的问题是, 亲和吸附剂成本很高不适合一 次性使用, 而所用活性碳和无机吸附剂具有副作用 (例如较强的蛋白吸附能力、 凝血因子失活、 血浆 APTT值大幅提高、 等等)。 、  In the prior art in which a solid phase medium is adsorbed to remove an unwanted virus inactivating agent, the organic solvent virus inactivating agent adsorption medium is an inorganic adsorbent (for example, a perlite filter plate containing perlite:), an affinity adsorbent (for example). C-18 oleophilic resin), activated carbon. A problem with the prior art is that the affinity adsorbent is expensive and not suitable for single use, and the activated carbon and inorganic adsorbent used have side effects (e.g., strong protein adsorption capacity, inactivation of coagulation factors, and substantial increase in plasma APTT value, and many more). ,
现有用固相介质吸附去除不需要的 毒灭活剂的技术中, 光敏剂病毒灭活 剂吸附介质分别为活性碳 (例如, PCT/US94/U227; US6159375; US08/347564)、 二 氧 化 硅 及 离 子 交 换 树 脂 (WO9103933) 、 大 孔 吸 附 树 脂 (poly styrene-divinylbenzene , acrylic ester polymers, US6348309)。 现有技术的问 题是,活性碳、二氧化硅及离子交换树脂具有副作用 (例如较强的蛋白吸附能力、 血浆 APTT值大幅提高、 等等), 而大孔吸附树脂容易脱落而污染生物液体, 也 可形成副作用。  In the prior art in which a solid phase medium is adsorbed to remove an unwanted toxic inactivating agent, the photosensitizer virus inactivating agent adsorption medium is activated carbon (for example, PCT/US94/U227; US6159375; US08/347564), and Ion exchange resin (WO9103933), poly styrene-divinylbenzene, acrylic ester polymers (US6348309). A problem with the prior art is that activated carbon, silica, and ion exchange resins have side effects (e.g., strong protein adsorption capacity, a large increase in plasma APTT value, etc.), while macroporous adsorption resins tend to fall off and contaminate biological fluids. Side effects can also be formed.
总之, 现有用于处理生物液体的、 包括病毒灭活处理的方法, 其中所用处 理系统中的固相介质多有副作用, 例如: 蛋白质构象变化与量的损失; 血小板 构象变化与量的损失; 红血球构象变化与量的损失、 杂质颗粒的引入、 等等。 在现有白细胞去除技术中, 也存在类似现象。 因而, 在目前条件下, 开发一种 既有有效功能(:例如对病毒灭活剂的吸附或 /和病毒灭活)、 又仅有最小化的副作 用的病毒灭活方法, 仍是科学技术追求的目标。 In summary, there are existing methods for treating biological fluids, including virus inactivation treatments, where Solid phase media in the system have many side effects, such as: protein conformational change and amount loss; platelet conformational change and amount loss; red blood cell conformational change and amount loss, introduction of impurity particles, and the like. A similar phenomenon exists in the existing leukocyte removal technology. Thus, under current conditions, the development of a virus inactivation method that has both effective functions (eg, adsorption of virus inactivating agents and/or virus inactivation) with minimal side effects is still the pursuit of science and technology. The goal.
发明内容 Summary of the invention
本发明的目的, 在于提供既有有效的病毒灭活功能、 或 /和有效的病毒灭活 剂去除功能, 又基本没有、 或仅有最小化的副作用的病毒灭活方法, 以及用于 本发明的方法的处理系统和装置。本发明中,表述"有效的病毒灭活功能"是指使 It is an object of the present invention to provide a virus inactivation method that has both an effective virus inactivating function, or/and an effective viral inactivating agent removal function, with substantially no or only minimal side effects, and is useful in the present invention. Method of processing systems and devices. In the present invention, the expression "effective virus inactivating function" means
99.9%以上的一种或多种模式病毒被灭活的功能; 表述 "有效的病毒灭活剂去除 功能"是指使 90%以上的一种或多种病毒灭活剂被除去的功能。 More than 99.9% of the functions of one or more model viruses are inactivated; the expression "effective virus inactivating agent removal function" means a function of removing more than 90% of one or more virus inactivating agents.
本发明的方法的基本技术方案为: 通过使用既有有效功能、 又仅有最小化 的副作用的固相介质, 来实现本发明的目的。该技术方案包括实施方案: 1.一般 而言,在固相介质上加入屯化剂可使其在病毒灭活中的副作用最小化; 2.对特定 的病毒灭活剂而言, 选择功能 /副作用比最大化的固相介质也可实现同一目的。  The basic technical solution of the method of the present invention is to achieve the object of the present invention by using a solid phase medium having both effective functions and only minimal side effects. The technical solution includes an embodiment: 1. In general, the addition of a oximation agent to a solid phase medium minimizes side effects in virus inactivation; 2. For a particular virus inactivating agent, a selection function/ Side effects can be achieved for the same purpose as the maximum solid phase medium.
本发 BJ的病毒灭活方法, 至少包括: Α).提供生物液体; Β).提供处理系统; 和 C).使所述生物液体与所述处理系统接触, 其中所述处理系统至少包含固相介 质和容纳所述固相介质的室, 其中所述固相介质至少包含: a). 病毒灭活剂吸附 介质; 和 /或 b).病毒灭活介质, 所述病毒灭活介质包含病毒灭活剂吸附介质和 固定在其上的病毒灭活剂,其中所述病毒灭活剂吸附介质包括: (a).钝化吸附介质, 所述钝化吸附介质至少含吸附介质和固定在吸附介质上的钝化剂;或 /和 (b).物理 吸附介质, 所述物理吸附介质包括: (i).用作光敏剂病毒灭活剂吸附介质的光敏剂 物理吸附纤维, 所述光敏剂物理吸附纤维含光敏剂病毒灭活剂的有机高分子物 理吸附剂; 和 (ii).用作有机溶剂病毒灭活剂吸附介质的有机溶剂物理吸附介质, 所述有机溶剂物理吸附介质含有机溶剂病毒灭活剂的有机高分子物理吸附剂。  The virus inactivation method of the present invention BJ, comprising at least: Α) providing a biological fluid; Β) providing a treatment system; and C) contacting the biological fluid with the treatment system, wherein the treatment system comprises at least a solid a phase medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, the virus inactivating medium comprising a virus An inactivating agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: (a) a passivation adsorption medium, the passivation adsorption medium containing at least an adsorption medium and being fixed in adsorption a passivating agent on the medium; or/and (b) a physical adsorption medium, the physical adsorption medium comprising: (i) a photosensitizer physical adsorbing fiber used as a photosensitizer virus inactivating agent adsorption medium, the photosensitizer Physically adsorbing fibers containing an organic polymer physical adsorbent of a photosensitizer virus inactivating agent; and (ii) an organic solvent physical adsorption medium used as an organic solvent virus inactivating agent adsorption medium, the organic solvent physical adsorption medium containing an organic solvent The organic polymer physical adsorbent drug inactivating agent.
本发明的处理系统, 其为本发明的上述病毒灭活方法中所述的处理系统。 所述处理系统至少包含固相介质和容纳所述固相介质的室, 其中所述固相介质 至少包含: a).病毒灭活剂吸附介质; 和 /或 b).病毒灭活介质, 所述病毒灭活介 质包含病毒灭活剂吸附介质和固定在其上的病毒灭活剂,其中所述病毒灭活剂吸 附介质包括: (a).钝化吸附介质,所述钝化吸附介质至少含吸附介质和固定在吸附 介质上的钝化剂; 或 /和 (b).物理吸附介质, 所述物理吸附介质包括: (i).用作光敏 剂病毒灭活剂吸附介质的光敏剂物理吸附纤维,所述光敏剂物理吸附纤维含光 敏剂病毒灭活剂的有机高分子物理吸附剂; 和 (ii).用作有机溶剂病毒灭活剂吸附 介质的有机溶剂物理吸附介质,所述有机溶剂物理吸附介质含有机溶剂病毒灭活 剂的有机高分子物理吸附剂。 The treatment system of the present invention is the treatment system described in the above virus inactivation method of the present invention. The processing system comprises at least a solid phase medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, The virus inactivating medium comprises a virus inactivating agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: (a) a passivation adsorption medium, the passivation adsorption medium being at least a passivating agent comprising an adsorbent medium and immobilized on the adsorbent medium; or/and (b) a physical adsorbent medium comprising: (i) a photosensitizer physics used as a photosensitizer virus inactivating agent adsorption medium Adsorbing fibers, the photosensitizer physically adsorbing an organic polymer physical adsorbent containing a photosensitizer virus inactivating agent; and (ii) using as an organic solvent virus inactivating agent An organic solvent physical adsorption medium of the medium, the organic solvent physical adsorption medium containing an organic solvent physical adsorbent of the organic solvent virus inactivating agent.
本发明的装置, 其至少含上述处理系统。  The device of the present invention comprises at least the above described processing system.
本发明的实施方案将说明, 通过本发明的方法、 处理系统或装置可以达到 本发明的目的。 要指出的是, 本发明的方法、 处理系统或装置除其副作用小的 优点外, 还特别适于固相介质一次性使用的情况。  Embodiments of the invention will illustrate that the objects of the invention can be attained by the method, processing system or apparatus of the invention. It is to be noted that the method, treatment system or device of the present invention is particularly suitable for the case where the solid phase medium is used at one time, in addition to the advantages of its small side effects.
具体实施方式 detailed description
本专业的技术人员应当知道, 以下使用的术语, 仅用以描述本发明的具体 模式, 本发明内容也许不限于下述解释的定义。  It should be understood by those skilled in the art that the terms used below are merely used to describe the specific modes of the present invention, and the present invention may not be limited to the definitions explained below.
在本发明中,术语"生物液体"是指含或可能含有生物物质的液体,例如血液 及血液成分、 血浆及血浆衍生物、 生物制品、 生物药物、 等等。 本发明中, 术 语"单人份血液组分"是指从同一个人身上一次或多次采集的血液或 /和从中分离 出来的组分, 包括单人份血液、 单人份血浆、 单人份血小板、 单人份红血球、 等等。  In the present invention, the term "biological liquid" means a liquid containing or possibly containing a biological substance such as blood and blood components, plasma and plasma derivatives, biological products, biopharmaceuticals, and the like. In the present invention, the term "single human blood component" means blood collected from one or more times from the same person or/and components separated therefrom, including single blood, single plasma, single serving. Platelets, single red blood cells, and more.
在本发明中,术语"有效减小病原体危害" 是指包括有效的病毒灭活或 /和白 细胞去除在内的一种处理过程; 术语"病毒灭活"是指通过加入病毒灭活剂 (例如 In the present invention, the term "effectively reducing pathogen harm" means a treatment process including effective virus inactivation or/and leukocyte removal; the term "viral inactivation" means by adding a virus inactivating agent (for example)
SD剂、 光敏剂病毒灭活剂、等等)或 /和物理能量 (例如热、光、等等), 使制备物 中至少部分可能存在的病毒的传染活性有效降低的过程;术语"白细胞去除"包括 通过对白细胞的吸附和尺寸过滤等机理进行的减小白细胞浓度的过程; 表述"有 效"对病毒灭活而言是指使 99,9%以上的一种或多种模式病毒被灭活、 对白细胞 去除而言是指使 90%以上的白细胞被除去。 SD agent, photosensitizer virus inactivating agent, etc.) or/and physical energy (eg heat, light, etc.), a process for effectively reducing the infectious activity of at least a portion of the virus present in the preparation; the term "white blood cell removal" "Includes a process for reducing leukocyte concentration by mechanisms such as adsorption and size filtration of leukocytes; the expression "effective" means, for virus inactivation, 99.9% or more of one or more model viruses are inactivated, For leukocyte removal, it means that more than 90% of white blood cells are removed.
在本发明中,术语"病毒灭活处理"是指使病毒灭活剂与生物液体接触直到生 物液体中的病毒灭活剂被基本除去的全过程。 病毒包括膜病毒或 /和非脂包膜病 毒。 病毒灭活处理含病毒灭活过程, 很多情况下还含病毒灭活剂去除过程。 在 生产过程中, 病毒灭活处理与其它处理是顺序进行或 /和同时进行的, 因而上述 病毒灭活处理中也可部分地经历或 /和伴随病毒灭活以外的其它处理 (:例如蛋白 质的纯化、 等等)。  In the present invention, the term "viral inactivation treatment" refers to the entire process of bringing a virus inactivating agent into contact with a biological fluid until the viral inactivating agent in the biological fluid is substantially removed. Viruses include membrane viruses or/and non-lipid envelope viruses. The virus inactivation treatment contains a virus inactivation process, and in many cases, a virus inactivating agent removal process. In the production process, the virus inactivation treatment and other treatments are performed sequentially or/and simultaneously, and thus the above virus inactivation treatment may also partially undergo or/and be accompanied by other treatments other than virus inactivation (eg, protein) Purification, etc.).
本发明中,术语"病毒灭活剂"是指任何对生物液体中可能存在的全部或部分 病毒具有病毒灭活功能、 但可能不限于病毒灭活功能的添加剂, 例如光敏剂病 毒灭活剂 /光照处理中的光敏剂病毒灭活剂、 SD处理中的有机溶剂病毒灭活剂或 有机溶剂病毒灭活剂 /去污剂、 等等。 在很多情况下病毒灭活剂除了病毒灭活功 能还可能有其它功能。 例如补骨脂素除了病毒灭活、 还对一些其它生物体 (例如 细菌)有灭活作用, 这时病毒灭活剂又是细菌灭活剂、等等。术语"病毒灭活剂衍 生物"是病毒灭活剂在病毒灭活处理中形成的衍生物, 例如光敏剂病毒灭活剂 / 光照处理中作为病毒灭活剂的亚甲蓝在光活化时产生的衍生物 (例如天青 A和 B)。 In the present invention, the term "viral inactivating agent" means any additive which has a virus inactivating function for all or part of viruses which may be present in a biological fluid, but may not be limited to a virus inactivating function, such as a photosensitizer virus inactivating agent/ Photosensitizer virus inactivating agent in light treatment, organic solvent virus inactivating agent in SD treatment or organic solvent virus inactivating agent/detergent, and the like. In many cases, the virus inactivating agent may have other functions besides the virus inactivating function. For example, psoralen is inactivated by viruses, but also inactivated by some other organisms (such as bacteria), in which case the virus inactivating agent is a bacterial inactivating agent, and the like. The term "viral inactivation agent" "Biological" is a derivative of a virus inactivating agent formed in a virus inactivation treatment, such as a photosensitizer virus inactivating agent/a derivative of methylene blue which is a virus inactivating agent during photoactivation (for example, azure) A and B).
本发明中, 有机溶剂病毒灭活包括以有机溶剂病毒灭活剂 (简称 S)或有机溶 剂病毒灭活剂和去污剂的组合(简称 SD)为病毒灭活剂而迸行的病毒灭活。 SD 处理包括美国纽约血液中心发明的 SD处理方法 (美国专利 US4540573)、 及后来 经他人开发的衍生方法 (例如, 只加入 S的灭病毒方法以及 S与酒精共用的病毒 灭活处理方法等)。本发明中, 术语"有机溶剂病毒灭活剂 (本发明中简称 S)"是指 在病毒灭活处理的特定条件下使用的有机溶剂。 这些特定条件包括但不限于: 竞争吸附 (例如, 在生物液体的环境中往往含有影响光敏剂吸附的组分, 例如人 血浆中的脂类物质); 反应动力学条件较不利 (例如, 液相病毒灭活时病毒灭活剂 的浓度一般不大于 l%(w/v)); 反应条件较保守 (:例如, 通常反应温度小于 38° C 而 pH在 4-10之间); 等等。 因而, 从技术方案的角度讲, 本发明中的有机溶剂 病毒灭活剂区别于一般意义上的有机溶剂。 有机溶剂病毒灭活剂包括任何用于 病毒灭活的溶剂, 例如磷酸三丁酯 (本发明中简称 TnBP)、 甲醛、 乙醚、 等等。 值得说明的是, 有机溶剂病毒灭活剂往往与去污剂共用。 本发明中, 去污剂包 括任何用于病毒灭活的去污剂, 例如吐温类去污剂 (例如 Tween-80)和 Triton 类去污剂 (例如 Triton X 100)。 In the present invention, the organic solvent virus inactivation comprises virus inactivation by a combination of an organic solvent virus inactivating agent (S) or an organic solvent virus inactivating agent and a detergent combination (SD) for the virus inactivating agent. . The SD treatment includes an SD treatment method invented by the New York Blood Center (US Pat. No. 4,540,573), and a derivative method developed by others (for example, a virus-in addition method in which only S is added, and a virus inactivation treatment method in which S is shared with alcohol, etc.). In the present invention, the term "organic solvent virus inactivating agent (abbreviated as S in the present invention)" means an organic solvent used under specific conditions of virus inactivation treatment. These specific conditions include, but are not limited to: competitive adsorption (eg, in biological fluid environments often contain components that affect the adsorption of photosensitizers, such as lipids in human plasma); reaction kinetic conditions are less favorable (eg, liquid phase) The concentration of the virus inactivating agent when the virus is inactivated is generally not more than 1% (w/v) ; the reaction conditions are relatively conservative (for example, the reaction temperature is usually less than 38 ° C and the pH is between 4 and 10); Therefore, from the viewpoint of the technical solution, the organic solvent virus inactivating agent in the present invention is distinguished from the organic solvent in the general sense. The organic solvent virus inactivating agent includes any solvent for virus inactivation, such as tributyl phosphate (abbreviated as TnBP in the present invention), formaldehyde, diethyl ether, and the like. It is worth noting that organic solvent virus inactivating agents are often shared with detergents. In the present invention, the detergent includes any detergent for virus inactivation, such as a Tween-type detergent (e.g., Tween-80) and a Triton-based detergent (e.g., Triton X 100).
本发明中, 光敏剂病毒灭活包括以光敏剂病毒灭活剂为病毒灭活剂而进行 的病毒灭活;术语"光敏剂病毒灭活剂",是指在病毒灭活处理的特定条件下使用 的光敏剂。 这些特定条件包括但不限于: 竞争吸附 (例如, 在生物液体的环境中 往往含有影响光敏剂吸附的组分, 例如人血桨中的脂类物质); 反应动力学条件 较不利 (例如, 液相病毒灭活时病毒灭活剂的浓度一般不大于 Ιμπιοΐ/l); 反应条 件较保守 (例如, 通常反应温度小于 38*^而 11在 6-10之间); 等等。 因而, 从 技术方案的角度讲, 本发明中的光敏剂病毒灭活剂区别于一般意义上的光敏剂。 从种类上讲, 光敏剂病毒灭活剂包括但不限于染料类光敏剂病毒灭活剂、 金丝 桃素(hypericin)和补骨脂素类光敏剂病毒灭活剂、 等等。通常, 这些光敏剂病 毒灭活剂在适当光照射下可有高得多的病毒灭活效率。 本发明中, 术语 "染料 类光敏剂病毒灭活剂 "是指可用作光敏剂病毒灭活剂进行病毒灭活 (例如光化学 病毒灭活)的染料类物质, 例如, 酞菁(美国专利 US5232844)、 假蓝宝石 (sapphyrin) (美国专利 US5041078)、 吩噻嗪染料 (Lambrecht et al., Vox Sang. 60, 207-213, (1991) 、 和花青染料 (美国专利 US4915683)、 等等; 术语 "补骨脂素 类光敏剂病毒灭活剂" 是指可用以进行光敏剂病毒灭活剂病毒灭活 (例如光化学 病毒灭活)的补骨脂素类物质(美国专利 US4169204、 4294822、 4328239 和 4727027), 包括补骨脂素 (英语 Psoralen^ 补骨脂素衍生物、 等等。 补骨脂素衍 生物的例子有 8-methoxyproralen 5-methoxy psoralen S59 psoralen^ trioxsalen-, aminomethyltrimethylpsorale 等等。 In the present invention, the photosensitizer virus inactivation comprises virus inactivation with a photosensitizer virus inactivating agent as a virus inactivating agent; the term "photosensitizer virus inactivating agent" means under specific conditions of virus inactivation treatment. The photosensitizer used. These specific conditions include, but are not limited to: competitive adsorption (eg, in biological fluid environments often contain components that affect the adsorption of photosensitizers, such as lipids in human blood plasma); reaction kinetic conditions are less favorable (eg, liquid The concentration of the virus inactivating agent when the phase virus is inactivated is generally not more than Ιμπιοΐ/l); the reaction conditions are relatively conservative (for example, usually the reaction temperature is less than 38*^ and 11 is between 6-10); Thus, from the viewpoint of the technical solution, the photosensitizer virus inactivating agent in the present invention is distinguished from the photosensitizer in the general sense. Typically, photosensitizer virus inactivating agents include, but are not limited to, dye photosensitizer virus inactivating agents, hypericin and psoralen photosensitizer virus inactivating agents, and the like. Generally, these photosensitizer virus inactivating agents can have much higher virus inactivation efficiencies under appropriate light exposure. In the present invention, the term "dye-based photosensitizer virus inactivating agent" means a dye-like substance which can be used as a photosensitizer virus inactivating agent for virus inactivation (for example, photochemical virus inactivation), for example, phthalocyanine (US Patent No. 5,232,844) ), sapphyrin (US Patent US5041078), phenothiazine dyes (Lambrecht et al., Vox Sang. 60, 207-213, (1991), and cyanine dyes (US Patent US 4,915,683), etc.; "Psoralen-type photosensitizer virus inactivating agent" means that the photosensitizer virus inactivating agent can be used for virus inactivation (eg photochemistry) Viral inactivated psoralen substances (U.S. Patents 4,169,204, 4,294,822, 4,328,239 and 4,727,027), including psoralen (English Psoralen^ psoralen derivatives, etc. Examples of psoralen derivatives) There are 8-methoxyproralen 5-methoxy psoralen S59 psoralen^ trioxsalen-, aminomethyltrimethylpsorale and so on.
本发明中, 术语 "副作用"是指在使用固相介质、 处理系统或装置时, 它们 除具有有用的功能、 也可能具有的引起生物液体生物活性的不需要的变化的能 力, 例如下述一种或多种变化: 蛋白质总量的减小、 对凝血处理系统的干扰、 对血小板形态的改变、 对溶血性的提高、 等等。 本发明中, 术语 "特异性吸附" 是指病毒灭活剂吸附介质对病毒灭活剂的吸附、 或去白细胞介质对白细胞的吸 附; 术语 "非特异性吸附"是指病毒灭活剂吸附介质对病毒灭活剂以外物质的吸 附、 或去白细胞介质对白细胞以外物质的吸附。 非特异性吸附包括对生物液体 的所需要的性质有益的吸附 (例如除脂)和不需要的吸附 (例如凝血因子活性的减 少)。 固相介质的不需要的反应活性包括不需要的吸附和不需要的其它反应活性 (例如催化活性、 酶激活活性、 参与生物液体中反应的活性、 等等)。  In the present invention, the term "side effects" refers to the ability of a solid phase medium, treatment system or device to have unwanted functions in addition to having a useful function, and may also cause an undesirable change in the biological activity of the biological fluid, such as the following One or more changes: a reduction in the total amount of protein, interference with the coagulation system, changes in platelet morphology, improvement in hemolytic properties, and the like. In the present invention, the term "specific adsorption" refers to adsorption of a virus inactivating agent to a virus inactivating agent, or removal of leukocyte media by leukocyte medium; the term "non-specific adsorption" refers to a virus inactivating agent adsorption medium pair Adsorption of substances other than viral inactivating agents, or removal of substances other than leukocytes by leukocyte mediators. Non-specific adsorption includes adsorption (e.g., degreasing) and unwanted adsorption (e.g., reduction in clotting factor activity) that are beneficial to the desired properties of the biological fluid. Unwanted reactivity of the solid phase medium includes unwanted adsorption and unwanted other reactive activities (e.g., catalytic activity, enzyme activating activity, activity involved in reactions in biological fluids, etc.).
在本发明中, 术语"固相介质", 亦简称介质, 是指具有某种功能 (例如吸附 功能、 病毒灭活功能、 等等)的固相材料; 术语"病毒灭活剂吸附介质", 有时简 称吸附介质, 是指至少具有、 但可能不限于对病毒灭活剂或 /和病毒灭活剂衍生 物有吸附功能 (例如有时它也可具有过滤功能、 除白细胞功能、等等:)、 且能将生 物液体中病毒灭活剂或 /和病毒灭活剂衍生物的浓度最终降至可接受的水平 (例 如磷酸三丁酯浓度小于 10ppm、 亚甲蓝浓度小于 0.03 g /ml、 等等)的固相材料, 吸附介质可以是纯物质 (例如活性炭粉、 木纤维、 等等)、 也可以是混合物 (例如 含纤维素的滤材、含活性碳的滤材、 C18层析胶、等等);术语"病毒灭活介质" 是 指至少具有、但可能不限于病毒灭活功能 (例如有时它也可具有过滤功能)的固相 介质, 例如病毒灭活剂 /吸附介质复合物,· 术语"去白细胞介质 "是指任何可用于 去白细胞的固相介质。 In the present invention, the term "solid phase medium", also referred to as medium, refers to a solid phase material having a certain function (for example, adsorption function, virus inactivation function, etc.); the term "virus inactivating agent adsorption medium", Sometimes referred to as an adsorption medium, it means having at least, but not limited to, a function of adsorbing a virus inactivating agent or/and a virus inactivating agent derivative (for example, sometimes it may also have a filtering function, a leukocyte function, etc.), And can ultimately reduce the concentration of viral inactivating agents or/and virus inactivating agent derivatives in biological fluids to acceptable levels (eg, tributyl phosphate concentration less than 10 ppm, methylene blue concentration less than 0.03 g / ml, etc. The solid phase material, the adsorption medium may be a pure substance (such as activated carbon powder, wood fiber, etc.), or may be a mixture (for example, a cellulose-containing filter medium, an activated carbon-containing filter medium, a C18 chromatography gel, etc.) The term "viral inactivating medium" means a solid phase medium having at least, but not necessarily limited to, a virus inactivating function (for example, it may also have a filtering function sometimes), such as a virus inactivating agent/adsorption medium complex, the term" Interleukin "refers to any solid phase media may be used to leukocytes.
在本发明中,术语"病毒灭活剂吸附剂"是指在病毒灭活处理的特定条件下通 过吸附有效除去病毒灭活剂的吸附剂。 这些特定条件包括: 特定的病毒灭活剂; 特定的吸附环境 (例如, 生物液体中往往含有影响吸附的组分, 例如人血浆中的 脂类物质); 特定的吸附动力学条件 (例如, 如本发明的实施例, 生物液体中病毒 灭活剂的浓度较小, 然而要求吸附后残存的病毒灭活剂的浓度极端低, 例如小 于 lOppm或小于 0.01μηιο1/1); 特定的吸附反应条件 (例如, 为保持生物液体的活 性, 吸附温度通常限于 0-37°C, 吸附 pH值通常限于 3.8-9.8); 等等。 因而, 从 技术方案的角度讲, 本发明中的病毒灭活剂吸附剂区别于一般意义上的吸附剂。 本发明中,术语"有机高分子化学吸附剂"是指通过在有机高分子上引入足够 量的生成化学键的成键基团 (例如离子基团、 配位基团、 等等)来制备、 因而主 要通过所引入的成键基团生成化学键 (例如离子键、配位键、等等)进 ¾^及附的试 剂,例如离子交换纤维、离子交换颗粒和螯合剂; "有机髙分子亲和吸附剂"是指 通过在有机髙分子上引入足够量的亲和配体 (例如肼的端基, 或病毒 RNA或 DNA的仿制品对某些光敏剂病毒灭活剂的亲和吸附)来制备、因而主要通过所引 入的亲和配体进行吸附的试剂, 例如亲和纤维、 亲和颗粒、 等等; "有机高分子 物理吸附剂"是指上述有机高分子化学吸附剂和有机高分子亲和吸附剂之外的 有机高分子吸附剂, 例如植物纤维、聚苯烯、等等,包括未进行专门的亲和基团 化或 /和未进行专门的离子交换基团化改性的有机高分子。 需要指出的是, 有机 高分子不包括活性碳。 本发明实施例中, 有机高分子物理吸附剂不包括目前己 知的有机高分子亲和吸附剂和有机高分子离子交换吸附剂, 这些功能化有机高 分子均经过专门的亲和基团化 (;例如亲和纤维、 亲和颗粒)或 /和专门的离子交换 基团化 (例如离子交换纤维、离子交换颗粒)改性。有机高分子物理吸附剂不必须 通过在有机高分子上引入足够量的离子交换基团或有机配体来制备、 因而主要 通过有机高分子上的弱相互作用 (包括范德华引力、 偶极-偶极相互作用、 氢健、 等等)和在制备过程中导入的少量 (端基、 或 /和链上 5%以下的取代基)离子基团 (例如植物纤维上的少量酸基)的作用进行吸附的试剂。 In the present invention, the term "viral inactivating agent adsorbent" means an adsorbent which is effective for removing a virus inactivating agent by adsorption under specific conditions of virus inactivation treatment. These specific conditions include: specific viral inactivating agents; specific adsorption environments (eg, biological fluids often contain components that affect adsorption, such as lipids in human plasma); specific adsorption kinetic conditions (eg, eg In the embodiment of the present invention, the concentration of the virus inactivating agent in the biological liquid is small, but the concentration of the virus inactivating agent remaining after the adsorption is required to be extremely low, for example, less than 10 ppm or less than 0.01 μηιο 1 / 1) ; specific adsorption reaction conditions ( For example, to maintain the activity of the biological fluid, the adsorption temperature is usually limited to 0-37 ° C, the adsorption pH is usually limited to 3.8-9.8); Thus, from the viewpoint of the technical solution, the virus inactivating agent adsorbent in the present invention is distinguished from the adsorbent in the general sense. In the present invention, the term "organic polymer chemical adsorbent" is prepared by introducing a sufficient amount of a bonding group (for example, an ionic group, a coordinating group, etc.) which generates a chemical bond to an organic polymer, thereby Mainly through the introduced bonding groups to generate chemical bonds (such as ionic bonds, coordination bonds, etc.) into the reagents, such as ion exchange fibers, ion exchange particles and chelating agents; "organic 髙 molecular affinity adsorption "agent" is prepared by introducing a sufficient amount of an affinity ligand (eg, a terminal group of hydrazine, or an affinity of a viral RNA or DNA for the affinity adsorption of certain photosensitizer virus inactivating agents) onto an organic hydrazine molecule, Therefore, the reagent mainly adsorbed by the introduced affinity ligand, such as affinity fiber, affinity particle, etc.; "organic polymer physical adsorbent" refers to the above organic polymer chemical adsorbent and organic polymer affinity Organic polymeric adsorbents other than adsorbents, such as plant fibers, polyphenylenes, and the like, include organic polymers that have not undergone specific affinity grouping or/and have not undergone special ion exchange group modification. . It should be noted that the organic polymer does not include activated carbon. In the embodiment of the present invention, the organic polymer physical adsorbent does not include the currently known organic polymer affinity adsorbent and the organic polymer ion exchange adsorbent, and these functionalized organic polymers are subjected to special affinity grouping ( Modifications such as affinity fibers, affinity particles) or/and specialized ion exchange groups (eg, ion exchange fibers, ion exchange particles). The organic polymer physical adsorbent does not have to be prepared by introducing a sufficient amount of ion exchange groups or organic ligands onto the organic polymer, and thus mainly through weak interactions on the organic polymer (including van der Waals attraction, dipole-dipole Interaction, hydrogenation, etc.) and adsorption of a small amount of (end group, or / and 5% or less substituents on the chain) ionic groups (for example, a small amount of acid groups on plant fibers) introduced during the preparation process Reagents.
本发明中, 术语"有机高分子物理吸附剂"中的有机高分子包括天然高分子, 例烈天然聚多糖。 在本发明实施例中, 所述聚多糖包括下述一种或多种物质: 纤维素、 葡聚糖、 琼脂糖、 壳聚糖、 淀粉。 纤维素衍生物的例子包括硝酸纤维 素、 醋酸纤维素、 甲基纤维素、 羧甲基纤维素、 等等。 尽管聚多糖作为载体己 被用于除病毒灭活剂中, 例如硝酸纤维素被用作光敏剂病毒灭活剂配体的载体 而用于除光敏剂病毒灭活剂和除白细胞滤器 (美国专利 US08/179567、08/204102、 08/347564)、含聚多糖的凝胶被作 SD剂吸附剂 (:例如 C18)的载体而用于除 SD剂、 等等, 本发明实施例的另一个意想不到的结果, 则是聚多糖本身对染料 (例如亚 甲蓝)的足够强的、 动力学稳定的可重复的吸附。 在本发明的实施例中, 某些滤 材 (例如 Seitz-BiolO、Seitz-Bio20、Seitz-bio40)、某些多孔颗粒 (例如 Sephadex G10、 Sephadex G25 Sephadex G50、 Sepharose )都含有聚多糖。  In the present invention, the organic polymer in the term "organic polymer physical adsorbent" includes a natural polymer, such as a strong natural polysaccharide. In an embodiment of the invention, the polysaccharide comprises one or more of the following: cellulose, dextran, agarose, chitosan, starch. Examples of the cellulose derivative include nitrocellulose, cellulose acetate, methyl cellulose, carboxymethyl cellulose, and the like. Although polyglycans have been used as carriers in addition to viral inactivating agents, for example, nitrocellulose has been used as a carrier for photosensitizer virus inactivating agent ligands in addition to photosensitizer virus inactivating agents and leukocyte-removing filters (US patent) US08/179567, 08/204102, 08/347564), a polysaccharide containing polypolysaccharide is used as a carrier for an SD agent adsorbent (for example, C18) for use in addition to an SD agent, etc., another meaning of the embodiment of the present invention Unexpected results are a sufficiently strong, kinetically stable, reproducible adsorption of the polysaccharide itself to the dye (e.g., methylene blue). In an embodiment of the invention, certain filters (e.g., Seitz-BiolO, Seitz-Bio20, Seitz-bio 40), certain porous particles (e.g., Sephadex G10, Sephadex G25 Sephadex G50, Sepharose) all contain a polysaccharide.
本发明中, 术语"有机高分子物理吸附剂"中的有机高分子包括合成高分子, 例如聚烯烃、 聚氨酯、 聚焼酯、 等等。 在本发明实施例中, 所述合成高分子包 括聚烯烃, 例如聚苯乙烯、 聚氯乙烯、 聚乙烯醇、 等等。 在本发明的实施例中, 某些滤材 (例如 Seitz-Supradurl00、 Seitz-Supradur200、 Seitz-Supradur 500)、 某些 多孔颗粒 (:例如交联丙烯酸大孔吸附树脂、 交联聚苯乙烯大孔吸附树脂、 聚苯乙 烯一聚丙烯腈大孔吸附树脂、 聚甲基丙烯酸甲酯树脂、 聚异丁烯等)都含有聚烯 烃。 In the present invention, the organic polymer in the term "organic polymer physical adsorbent" includes synthetic polymers such as polyolefins, polyurethanes, polydecyl esters, and the like. In the embodiment of the present invention, the synthetic polymer includes a polyolefin such as polystyrene, polyvinyl chloride, polyvinyl alcohol, or the like. In an embodiment of the invention, certain filters (eg Seitz-Supradurl00, Seitz-Supradur200, Seitz-Supradur 500), certain Porous particles (for example, crosslinked acrylic macroporous adsorption resin, crosslinked polystyrene macroporous adsorption resin, polystyrene-polyacrylonitrile macroporous resin, polymethyl methacrylate resin, polyisobutylene, etc.) all contain poly Olefins.
本发明中, 术语"钝化剂,,是指对固相介质或处理系统的其它组成物上的不 需要的反应活性 (:例如副作用)具有钝化作用、但可能不限于钝化作用的试剂。本 发明中, 术语"复合物" 是指两种或两种以上的组成通过吸附或其它物理化学、 化学、或生物化学作用联接形成的材料。例如,术语"钝化剂 /吸附介质复合物"是 指最少组成为病毒灭活剂的吸附介质、 和吸附介质的钝化剂的复合物; 术语"病 毒灭活剂 /吸附介质 /钝化剂复合物" 是指最少组成为病毒灭活剂、病毒灭活剂的 吸附介质、 和吸附介质的钝化剂的复合物。  In the present invention, the term "passivating agent" refers to a reagent which has a passivation effect on an undesired reaction (eg, side effects) on a solid phase medium or other composition of a treatment system, but may not be limited to passivation. In the present invention, the term "composite" means a material in which two or more compositions are joined by adsorption or other physicochemical, chemical, or biochemical interaction. For example, the term "passivation agent/adsorption medium complex" "refers to a complex of at least an adsorbent medium composed of a virus inactivating agent and a passivating agent of an adsorbent medium; the term "viral inactivating agent / adsorbent medium / passivator complex" means a minimum composition of a virus inactivating agent a complex of an adsorption medium for the virus inactivating agent and a passivating agent for the adsorption medium.
在本发明中, 处理系统可独立作为一个装置 (例如滤器、 层析柱、 等等)、 或 作为装置的一个构成 (例如单人份血液或组分处理装置中的滤器、层析柱、等等)。 在本发明中, 术语 "装置"是指最小组成物为上述处理系统的、 可实际应用于 处理生物液体的结构, 例如滤器、 层析柱、 试剂盒 (英语 Kit:)、 等等; 术语 "室" 指一种至少可容纳固相介质的中空容器, 例如滤器的中空滤筒、 层析柱的中空 柱、 等等。  In the present invention, the processing system can be used as a device (eg, a filter, a chromatography column, etc.), or as a component of the device (eg, a filter, column, etc. in a single blood or component processing device). Wait). In the present invention, the term "device" means a structure in which the minimum composition is the above-described treatment system, which can be practically applied to the treatment of biological fluids, such as filters, chromatography columns, kits (English Kit:), etc.; "Case" means a hollow vessel that can hold at least a solid phase medium, such as a hollow filter cartridge of a filter, a hollow column of a chromatography column, and the like.
本发明中,术语"滤材"一般地是指可仅允许一定尺寸之下的结构流过的固相 介质。 因而, 本发明中的所述滤材往往用作固-液相反应中的固定相, 例如对病 毒灭活剂有吸附功能的滤材 (简称特异性吸附滤材)。一种滤材是否特异性吸附滤 材, 除了生产厂家提供的信息 (例如活性炭滤板),更主要是看它是否在病毒灭活 处理中是否有特异性吸附功能。 例如, 通常认为无吸附功能的 Seitz-Bio滤板便 在本发明的实施例中被发现是一种染料类光敏剂病毒灭活剂吸附介质。 滤材包 括绝对过滤滤材和深层过滤滤材。 本发明中滤材的一个优选方案是深层过滤滤 材。 尽管不拟进入理论讨论, 但深层过滤滤材中流体流径较长, 对其功能 (例如 吸附、病毒灭活)实现具有重要影响。滤材有多种形式, 例如滤膜、滤板、滤棒、 滤芯、 滤毡、 等等。  In the present invention, the term "filter material" generally means a solid phase medium which can only allow a structure below a certain size to flow. Therefore, the filter medium in the present invention is often used as a stationary phase in a solid-liquid phase reaction, for example, a filter material having an adsorption function for a virus inactivating agent (referred to as a specific adsorption filter medium). Whether a filter material specifically adsorbs the filter material, in addition to the information provided by the manufacturer (such as activated carbon filter plate), it is mainly to see if it has a specific adsorption function in the virus inactivation treatment. For example, a Seitz-Bio filter plate which is generally considered to have no adsorption function is found to be a dye-based photosensitizer virus inactivating agent adsorption medium in the examples of the present invention. The filter media includes absolute filter media and depth filter media. A preferred embodiment of the filter material of the present invention is a depth filtration filter. Although not intended to be discussed in theory, the fluid flow path in the deep filter media is long and has a significant impact on its function (eg adsorption, virus inactivation). Filter media come in many forms, such as filters, filter plates, filter rods, filter cartridges, filter mats, and more.
本发明中,术语"滤器"是指至少包含滤材及其持有物的装置,其包括但不限 于公知的滤器。 所述持有物包括将所述固相介质周边用压力封闭的结构, 例如 滤盘、 滤筒、 滤柱、 等等。 本发明的以下实施例的滤器中, 所述持有物一般地 包括: 容纳所述固相介质的容器、 固定所述固相介质的上端结构和下端结构、 及固相介质周边用压力封闭滤材式固相介质的结构 (例如, 由于在上端结构和下 端结构上高出的凸体在封闭时对固相介质周边的接触力而使固相介质封闭以避 免液体泄漏的压力封闭结构)。 此外, 还可增加其它功能结构, 例如在上端结构 或 /和下端结构上引入有特殊功能的结构 (例如除菌滤膜、 等等)。 本专业的技术 人员应当知道, 使用本发明的以下实施例的滤器式装置, 通过与不同的结构组 合, 可制备出很多具有不同用处的滤器。 In the present invention, the term "filter" means a device comprising at least a filter material and its holder, including but not limited to known filters. The holder includes a structure that seals the periphery of the solid phase medium with pressure, such as a filter disc, a filter cartridge, a filter column, and the like. In the filter of the following embodiments of the present invention, the holder generally includes: a container for accommodating the solid phase medium, an upper end structure and a lower end structure for fixing the solid phase medium, and a pressure sealing filter for surrounding the solid phase medium The structure of the solid-state medium of the material (for example, a pressure-closed structure in which the solid phase medium is closed to avoid liquid leakage due to the contact force of the convex body which is higher on the upper end structure and the lower end structure to the periphery of the solid phase medium when closed). In addition, other functional structures can be added, such as in the upper structure. Or a structure with a special function introduced on the lower end structure (for example, a sterile filter membrane, etc.). Those skilled in the art will appreciate that a plurality of filters having different uses can be prepared by using a filter device of the following embodiments of the present invention by combining with different structures.
本发明中, 术语"染座"是指可结合染料的结构,包括物理结构、物理化学结 构、 化学结构 (例如基团)、 等等; 术语"纤维"是指长度 /直径比大于 10、 且平均 直径小于 1mm的材料; 术语"纤维微结构" (英语 Fibrets)是指纤维经专门的生产 工艺或专门的加工形成的一种具有很多的不规则的微分枝和很大的表面积的纤 维的微结构, 这种纤维微结构非常短 (小于 1mm)且非常细 (小于 50μιη), 例如 Seitz-BiolO, Seitz-Bio20、 Seitz-bio40滤板就含纤维微结构。  In the present invention, the term "dyeing" means a structure which can bind a dye, including a physical structure, a physicochemical structure, a chemical structure (for example, a group), and the like; the term "fiber" means a length/diameter ratio of more than 10, and A material having an average diameter of less than 1 mm; the term "Fibrets" refers to a fiber having a large number of irregular micro branches and a large surface area formed by a special production process or special processing. Structure, this fiber microstructure is very short (less than 1 mm) and very fine (less than 50 μm), for example Seitz-BiolO, Seitz-Bio20, Seitz-bio40 filter plates contain fiber microstructure.
本发明的第一个方面涉及本发明的病毒灭活方法。  A first aspect of the invention relates to a virus inactivation method of the invention.
本发明的病毒灭活方法, 至少包括: A).提供生物液体; B).提供处理系统; 和 C).使所述生物液体与所述处理系统接触, 其中所述处理系统至少包含固相介 质和容纳所述固相介质的室, 其中所述固相介质至少包含: a). 病毒灭活剂吸附 介质; 和 /或 b).病毒灭活介质, 所述病毒灭活介质包含病毒灭活剂吸附介质和 固定在其上的病毒灭活剂,其中所述病毒灭活剂吸附介质包括 :0).钝化吸附介质, 所述钝化吸附介质至少含吸附介质和固定在吸附介质上的钝化剂;或 /和 (b).物理 吸附介质, 所述物理吸附介质包括 :(i).用作光敏剂病毒灭活剂吸附介质的光敏剂 物理吸附纤维,所述光敏剂物理吸附纤维含光敏剂病毒灭活剂的有机髙分子物 理吸附剂; 和 (ii).用作有机溶剂病毒灭活剂吸附介质的有机溶剂物理吸附介质, 所述有机溶剂物理吸附介质含有机溶剂病毒灭活剂的有机高分子物理吸附剂。  The virus inactivating method of the present invention comprises at least: A) providing a biological fluid; B) providing a treatment system; and C) contacting the biological fluid with the treatment system, wherein the treatment system comprises at least a solid phase a medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, the virus inactivating medium comprising a virus a living agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: 0) a passivation adsorption medium, the passivation adsorption medium containing at least an adsorption medium and being fixed on the adsorption medium a passivating agent; or/and (b) a physical adsorbing medium comprising: (i) a photosensitizer physic adsorbing fiber used as a photosensitizer virus inactivating agent adsorption medium, said photosensitizer physically adsorbing An organic hydrazine molecular physical adsorbent containing a photosensitizer virus inactivating agent; and (ii) an organic solvent physical adsorption medium used as an organic solvent virus inactivating agent adsorption medium, the organic solvent physical adsorption medium containing organic solvent The organic polymer physical adsorbent inactivating agent.
本发明的方法基于本发明实施例中出人意料的结果:  The method of the present invention is based on unexpected results in embodiments of the invention:
1).关于纯化吸附介质的应用:  1). Application for purification of adsorption media:
由于生物液体产品标准中病毒灭活剂的许可残留值通常很低 (例如欧洲药典 中 TnBP许可残留值小于 10ppm), 通常的看法是: 对吸附介质是不宜钝化的。 令人惊奇的是, 本发明实施例中一些吸附介质固定钝化剂形成吸附介质 /钝化剂 复合物之后, 仍然可用于吸附除出病毒灭活剂, 而且与吸附介质比较具有更小 副作用。  Since the permissible residual value of the virus inactivating agent in the bio-liquid product standard is usually very low (for example, the TnBP approved residue value in the European Pharmacopoeia is less than 10 ppm), the general view is: It is not suitable for passivation of the adsorption medium. Surprisingly, in some embodiments of the present invention, after the adsorption medium is fixed to form the adsorption medium/passivator compound, the adsorption medium can still be used for adsorption removal of the virus inactivating agent, and has less side effects than the adsorption medium.
此外, 由于对病毒灭活的要求很高 (通常要求病毒滴度下降 103以上), 通常 的看法是:对病毒灭活介质 (特别是病毒灭活剂不在长链吸附基团上的固相介质, 例如 SD-活性炭)进行钝化可能降低病毒灭活介质的灭活效率。 因而, 在病毒灭 活介质的开发中均未引入钝化处理。 然而, 本发明的实施例将说明, 使用含钝 化剂的病毒灭活介质, 既有效地保护了固相介质所接触的液相介质的生物活性, 又有效地进行了病毒灭活, 从而达到了本发明的目的。 此外, 去白细胞介质通常具有对白细胞或多或少的吸附能力, 通常的观念 会认为加入钝化剂虽可抑制其副作用, 但也可能降低其对白细胞的吸附作用。 然而令人吃惊的是, 在本发明的一个实施例中, 加入钝化剂抑制了其副作用, 但仍可用其有效去除白细胞。 在一个优选方案中, 本发明的第一种处理系统可 用于同时去除病毒灭活剂 (或 /和病毒灭活剂衍生物)和白细胞。 In addition, due to the high requirement for virus inactivation (usually requiring a virus titer to drop by more than 10 3 ), the general view is that the virus is inactivated medium (especially the solid phase of the virus inactivating agent not on the long-chain adsorption group) Passivation of a medium, such as SD-activated carbon, may reduce the inactivation efficiency of the virus inactivating medium. Thus, no passivation treatment was introduced in the development of the virus inactivating medium. However, embodiments of the present invention will demonstrate that the use of a virus inactivating medium containing a passivating agent not only effectively protects the biological activity of the liquid medium contacted by the solid phase medium, but also effectively inactivates the virus, thereby achieving The object of the invention. In addition, the leukocyte-removing medium usually has more or less adsorption capacity to white blood cells. The general idea is that the addition of a passivating agent can inhibit its side effects, but it may also reduce its adsorption on white blood cells. Surprisingly, however, in one embodiment of the invention, the addition of a passivating agent inhibits its side effects, but it can still be used to effectively remove leukocytes. In a preferred embodiment, the first treatment system of the invention can be used to simultaneously remove viral inactivating agents (or/and viral inactivating agent derivatives) and white blood cells.
2).关于物理吸附介质的应用:  2). About the application of physical adsorption media:
一般地说, 通过含吸附剂的固相介质除去添加剂是一种久以使用的公知技 术。 本专业的技术人员应当知道, 通过含病毒灭活剂吸附剂的固相介质可除去 病毒灭活剂。 由于具吸附功能的固相介质一般地说是非常之多, 而病毒灭活剂 包括的种类亦不少, 因而开发出含对特定病毒灭活剂具有效吸附功能的病毒灭 活剂吸附介质的特定处理系统一直是很艰难的创造性工作, 开发出具特定的病 毒灭活剂的有效吸附、 而又基本上没有有意义的非特异性吸附 (例如, 基本上无 副作用:)的处理系统则是更加艰难的创造性工作。  In general, the removal of additives by solid phase media containing adsorbents is a well-known technique that has long been used. Those skilled in the art will appreciate that the viral inactivating agent can be removed by a solid phase medium containing a viral inactivating agent adsorbent. Since the solid phase medium with adsorption function is generally very much, and the virus inactivating agent includes many kinds, the virus inactivating agent adsorption medium containing the adsorption function for the specific virus inactivating agent is developed. Certain processing systems have been a very difficult creative task, and it has become more difficult to develop a treatment system that has a specific adsorption of a specific viral inactivating agent without substantial non-specific adsorption (for example, essentially no side effects:). Creative work.
关于光敏剂物理吸附纤维的应用, 也基于本发明实施例的意想不到的结果。 例如,尽管: A)某些染料 (:例如亚甲蓝)是人血液或人血液成分中最常用的病毒灭 活剂之一; B)含有染座的纤维 (例如天然纤维)可吸附染料; C)甚至含有天然纤维 的滤材已经长期用于生物制品生产, 然而迄今为止未见将天然纤维直接用作染 料类光敏剂病毒灭活剂的吸附介质。 这或许是熟识无睹, 或许是对其吸附能力 能否满足在浓度较低 (通常小于 1μπι01/ιη1)、温度不高 (通常小于 36Ό)的条件下染 料类光敏剂病毒灭活剂的有效吸附缺乏信心。 然而, 根据本发明的实施例, 今 人吃惊的是: 利用天然纤维或非离子交换纤维的合成纤维来进行染料病毒灭活 剂的有效去除, 并不比利用染料来对纤维进行染色更难、 甚至更易实现。 例如, 亚甲蓝通常不用作天然纤维的染色, 但利用天然纤维却可有效去除亚甲蓝病毒 灭活剂。 根据本发明的实施例, 这些纤维不仅对染料类光敏剂病毒灭活剂有吸 附能力、 且具有特征意义的是: 其吸附是足够强的 (例如吸附后亚甲蓝的浓度降 至 0.03 g /ml以下); 其吸附是动力学稳定的 (例如在生物液体不断流过滤板时其 上吸附的亚甲蓝不出现有意义的解吸); 其吸附是可重复的 (例如, 在同样条件下 其吸附重现性为 100%); 和其吸附是足够特异的 (例如, 基本上无副作用)。 The use of photosensitizers for physically adsorbing fibers is also based on the unexpected results of embodiments of the present invention. For example, although: A) certain dyes (eg, methylene blue) are one of the most commonly used viral inactivating agents in human blood or human blood components; B) fibers containing dyed seats (eg, natural fibers) can adsorb dyes; C) Even filter media containing natural fibers have long been used for the production of biological products, but no adsorption medium for directly using natural fibers as a dye-based photosensitizer virus inactivating agent has been seen so far. This may be familiar, whether it is capable of satisfying the dye-based photosensitizer virus inactivating agent at a lower concentration (usually less than 1μπι 0 1/ιη1) and a low temperature (usually less than 36Ό). Effective adsorption lacks confidence. However, according to an embodiment of the present invention, it is surprising that the use of synthetic fibers of natural fibers or non-ion exchange fibers for the effective removal of dye virus inactivating agents is no more difficult and even more difficult than dyeing fibers. Easier to implement. For example, methylene blue is generally not used for dyeing natural fibers, but natural fibers are used to effectively remove methylene blue virus inactivating agents. According to an embodiment of the present invention, these fibers are not only adsorbable to the dye-based photosensitizer virus inactivating agent, but also have the characteristic that the adsorption is sufficiently strong (for example, the concentration of methylene blue after adsorption is reduced to 0.03 g / Mol is below); its adsorption is kinetically stable (for example, there is no significant desorption of methylene blue adsorbed on the filter plate when the biological liquid is continuously flowing); its adsorption is reproducible (for example, under the same conditions) The adsorption reproducibility is 100%); and its adsorption is sufficiently specific (for example, substantially no side effects).
本发明不拟进入理论讨论, 仅以含有机高分子物理吸附剂的纤维为例提供 一些观察。 也许由于下述原因, 有机高分子物理吸附剂的弱相互作用 (例如生产 过程中引入的微量离子基团、范德华引力、偶极-偶极相互作用、 氢健)就足够强 去有效吸附染料类光敏剂病毒灭活剂: 1).染料类光敏剂病毒灭活剂的 "上染 (dye up-take)"通常是以纤维为固定相、含染料类光敏剂病毒灭活剂的生物液体为流动 相在优选的固 -液相反应条件下进行的; 2)去除染料类光敏剂病毒灭活剂并不要 求均染; 3).就所用纤维的数量而言, 待吸附的染料类光敏剂病毒灭活剂的总量 不是很大; 4). 纤维的结构复杂性; 等等。 总之, 有机高分子物理吸附剂的这些 弱相互作用, 其对特定条件下的特定病毒灭活剂的是吸附是足够强的, 而对特 定条件下的生物液体的副作用又是足够小的, 因而具有相对高的特异性吸附能 力, 从而能实现本发明的目的。 The present invention is not intended to be discussed in theory, and only some of the fibers containing organic polymeric physical adsorbents are provided for some observations. Perhaps due to the following reasons, weak interactions of organic polymeric physical adsorbents (such as trace ionic groups introduced during production, van der Waals attraction, dipole-dipole interaction, hydrogen chemistry) are strong enough to effectively adsorb dyes Photosensitizer virus inactivating agent: 1). Dye-based photosensitizer The "dye up-take" of the virus inactivating agent is usually a biological liquid containing a fiber as a stationary phase and a dye-based photosensitizer virus inactivating agent. Flow The phase is carried out under the conditions of the preferred solid-liquid phase reaction; 2) the dye-based photosensitizer virus inactivating agent is not required to be uniformly dyed; 3) the dye-based photosensitizer virus to be adsorbed in terms of the amount of fibers used The total amount of inactivating agent is not very large; 4 ). The structural complexity of the fiber; In summary, these weak interactions of organic polymeric physical adsorbents are sufficiently strong for specific viral inactivating agents under specific conditions, and the side effects of biological fluids under specific conditions are small enough. It has a relatively high specific adsorption capacity, so that the object of the present invention can be achieved.
关于有机溶剂物理吸附介质的应用, 也基于本发明实施例的意想不到的结 果。 例如,尽管: A)有机溶剂病毒灭活剂 (例如 TnBP)是人血液或人血液成分中 最常用的病毒灭活剂之一; B)含聚烯烃的多孔颗粒可去除油污; C)甚至含聚烯 烃的多孔颗粒已经用于染料类病毒灭活剂的去除, 然而迄今为止未见将含聚烯 烃的多孔颗粒直接用作有机溶剂病毒灭活剂的吸附介质、 特别是用以同有机溶 剂病毒灭活剂结合制备病毒灭活介质。 与上述光敏剂病毒灭活剂的情况相似, 这或许是熟识无睹, 或许是对其吸附能力能否满足在浓度较低 (通常小于 1% w/v)、温度不高 (通常小于 37°C)的条件下有机溶剂病毒灭活剂的有效吸附缺乏信 心。 然而, 根据本发明的实施例, 这些多孔颗粒不仅对有机溶剂病毒灭活剂有 吸附能力、 且具有特征意义的是: 其吸附是足够强的 (例如吸附后 TnBP的浓度 降至 lOppm以下); 其吸附是动力学稳定的 (例如不出现有意义的 TnBP解吸); 其吸附是可重复的 (例如, 在同样条件下其吸附重现性为 100%); 和其吸附是足 够特异的 (例如, 基本上无副作用)。  The use of organic solvent physical adsorption media is also based on the unexpected results of embodiments of the present invention. For example, although: A) an organic solvent virus inactivating agent (eg TnBP) is one of the most commonly used viral inactivating agents in human blood or human blood components; B) porous particles containing polyolefin can remove oil stains; C) even containing Porous particles of polyolefin have been used for the removal of dye-type virus inactivating agents, however, no porous particles containing polyolefin have been used as an adsorption medium for organic solvent virus inactivating agents, especially for organic solvent viruses. The inactivating agent is combined to prepare a virus inactivating medium. Similar to the photosensitizer virus inactivating agent described above, this may be familiar, perhaps because its adsorption capacity can be met at lower concentrations (usually less than 1% w/v) and at low temperatures (usually less than 37°). Under the conditions of C), the effective adsorption of the organic solvent virus inactivating agent lacks confidence. However, according to an embodiment of the present invention, the porous particles not only have an adsorption capacity to the organic solvent virus inactivating agent, but also have the characteristic that: the adsorption is sufficiently strong (for example, the concentration of TnBP after adsorption is reduced to less than 10 ppm); The adsorption is kinetically stable (eg, no significant TnBP desorption occurs); its adsorption is reproducible (eg, its adsorption reproducibility is 100% under the same conditions); and its adsorption is sufficiently specific (eg , basically no side effects).
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包括有机溶剂病 毒灭活剂吸附介质, 且所述有机溶剂病毒灭活剂吸附介质至少含所述钝化吸附 介质, 或 /和有机溶剂物理吸附介质。  In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises an organic solvent virus inactivating agent adsorption medium, and the organic solvent virus inactivating agent adsorption medium contains at least the passivation adsorption medium. Or / and organic solvent physical adsorption medium.
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包括有机溶剂病 毒灭活介质, 且所述有机溶剂病毒灭活介质至少含有机溶剂病毒灭活剂、 和所 述钝化吸附介质、 或 /和有机溶剂物理吸附介质。  In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises an organic solvent virus inactivating medium, and the organic solvent virus inactivating medium contains at least an organic solvent virus inactivating agent, and the blunt The adsorption medium, or/and the organic solvent physical adsorption medium.
在本发明的病毒灭活方法的一项实施方案中, 所述有机溶剂物理吸附介质 包括大孔吸附树脂或 /和纤维。在本发明的实施例中,所用纤维 (有机溶剂物理吸附 纤维)含有机溶剂病毒灭活剂的有机高分子物理吸附剂,例如天然纤维 (例如 Seitz Bio系列滤板)、聚烯烃纤维 (例如 Seitz Supradur 80、 Seitz Supradur500、 Seitz Eco 1000、 等等)、 等等。 所用大孔吸附树脂 (有机溶剂物理吸附树脂)包括聚苯乙烯 颗粒 (例如 Amberlite XAD- 7HP、 Amberlite AD-16 ), 酚醛缩聚型颗粒、 聚丙 烯酸酯颗粒和含聚烷酯大孔吸附树脂。 除此之外, 所述有机溶剂物理吸附介质 还包括内表面含所述有机高分子的多孔非颗粒材料。 例如, 聚烯烃包被陶瓷、 等等。 In an embodiment of the virus inactivation method of the present invention, the organic solvent physical adsorption medium comprises a macroporous adsorption resin or/and a fiber. In the embodiment of the present invention, the fiber (organic solvent physical adsorption fiber) contains an organic polymer physical adsorbent containing an organic solvent virus inactivating agent, such as natural fiber (for example, Seitz Bio series filter plate), polyolefin fiber (for example, Seitz). Supradur 80, Seitz Supradur500, Seitz Eco 1000, etc.), etc. The macroporous adsorption resin (organic solvent physical adsorption resin) used includes polystyrene particles (for example, Amberlite XAD-7HP, Amberlite AD-16), phenolic polycondensation type particles, polyacrylate particles, and polyalkyl ester-containing macroporous adsorption resin. In addition to the above, the organic solvent physical adsorption medium further includes a porous non-particulate material having an inner surface containing the organic polymer. For example, polyolefin coated ceramics, and many more.
在本发明的病毒灭活方法的一项实施方案中, 所述有机溶剂病毒灭活剂的有 机高分子物理吸附剂包括聚烯烃。 例如上述聚烯烃纤维、 聚苯乙烯颗粒、 聚丙 烯酸酯颗粒。 在一项实施方案的一项优选方案中, 所述聚烯烃包括聚苯乙烯,尤 其是含聚苯乙烯的大孔吸附树脂。  In an embodiment of the virus inactivation method of the present invention, the organic polymer physical adsorbent of the organic solvent virus inactivating agent comprises a polyolefin. For example, the above polyolefin fibers, polystyrene particles, and polyacrylate particles. In a preferred embodiment of an embodiment, the polyolefin comprises polystyrene, especially a macroporous adsorption resin comprising polystyrene.
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包括光敏剂病毒 灭活剂吸附介质, 且所述光敏剂病毒灭活剂吸附介质至少含所述钝化吸附介质, 或 /和光敏剂物理吸附纤维。在本发明的实施例中,所用光敏剂物理吸附纤维包括 天然纤维、 纤维素基衍生纤维、 非功能化合成纤维。  In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises a photosensitizer virus inactivating agent adsorption medium, and the photosensitizer virus inactivating agent adsorption medium contains at least the passivation adsorption medium. Or / and the photosensitizer physically adsorbs the fibers. In the examples of the present invention, the photosensitizer physical adsorbing fibers used include natural fibers, cellulose-based derived fibers, and non-functionalized synthetic fibers.
在本发明的病毒灭活方法的一项实施方案中, 所述光敏剂物理吸附纤维包括 天然纤维。在本发明的实施例中,所用天然纤维包括植物纤维、 动物纤维 (例如蚕 丝)。 所用植物纤维包括棉纤维 (:例如无脂棉)、 木纤维 (在滤板中) 、 麻纤维、 纸 浆 (例如纸)。 所述纤维还包括纤维素基衍生纤维 (例如, 粘胶纤维、 醋酸纤维、 等等)。在本发明的病毒灭活方法的一项实施方案中,所述天然纤维包括木纤维。  In an embodiment of the virus inactivation method of the present invention, the photosensitizer physically adsorbing fibers comprise natural fibers. In the examples of the present invention, the natural fibers used include plant fibers, animal fibers (e.g., silk). The plant fibers used include cotton fibers (e.g., non-fat cotton), wood fibers (in filter plates), hemp fibers, pulp (e.g., paper). The fibers also include cellulose based derived fibers (e.g., viscose, acetate, and the like). In an embodiment of the virus inactivation method of the invention, the natural fibers comprise wood fibers.
在本发明的病毒灭活方法的一项实施方案中, 所述光敏剂物理吸附纤维包 括合成纤维,优选非功能化合成纤维。在本发明的实施例中,所用非功能化合成纤 维包括亲和纤维和离子交换纤维以外的合成纤维,例如聚酯纤维、聚丙烯纤维 (例 如超细聚丙烯吸油纤维)、 聚酰氨纤维、 聚氨酯纤维和其它聚烯烃纤维。 在一项 优选方案中, 所述纤维具有下述一种或多种特征: A).平均直径 0.01-20μπι; Β). 卷曲度 1-2,· C).吸水量 25-50g/100g纤维。 此外, 众所周知, 纤维的其它结构参 数, 例如高分子的玻璃化温度、 纤维的动电层电位和等电点、 等等, 也对其吸 附能力构成影响。 In an embodiment of the virus inactivation method of the present invention, the photosensitizer physically adsorbing fibers comprise synthetic fibers, preferably non-functionalized synthetic fibers. In an embodiment of the invention, the non-functionalized synthetic fibers used include synthetic fibers other than affinity fibers and ion exchange fibers, such as polyester fibers, polypropylene fibers (eg, ultrafine polypropylene oil absorbing fibers), polyamide fibers, Polyurethane fibers and other polyolefin fibers. In a preferred embodiment, the fiber has one or more of the following characteristics: A). Average diameter 0.01-20 μπι ; Β). Curl degree 1-2, · C). Water absorption 25-50 g /100 g fiber. In addition, it is well known that other structural parameters of the fiber, such as the glass transition temperature of the polymer, the electrokinic layer potential of the fiber, and the isoelectric point, etc., also affect its adsorption capacity.
在本发明的病毒灭活方法的一项实施方案中, 所述合成纤维包括聚烯烃纤 维。 在本发明的实施例中,某些用作光敏剂物理吸附介质的滤材(例如 Seitz-SupradurlOO、 Seitz-Supradur200 Seitz-Supradur 500)都含有聚烯烃纤维。  In an embodiment of the virus inactivation method of the present invention, the synthetic fiber comprises a polyolefin fiber. In an embodiment of the invention, certain filter media (e.g., Seitz-SupradurlOO, Seitz-Supradur 200 Seitz-Supradur 500) used as a sensitizing agent for the photosensitizer contain polyolefin fibers.
在本发明的病毒灭活方法的一项实施方案中, 所述病毒灭活剂吸附介质还 另含有颗粒, 且所述颗粒含光敏剂病毒灭活剂的有机高分子物理吸附剂。 例如 下述处理系统: 在一个室中, 光敏剂病毒灭活剂吸附介质包括含苯乙烯的有机 高分子颗粒和位于颗粒下方的光敏剂物理吸附纤维 (例如聚烯烃纤维)的无纺布 或滤材。  In an embodiment of the virus inactivation method of the present invention, the virus inactivating agent adsorption medium further comprises particles, and the particles comprise an organic polymer physical adsorbent of a photosensitizer virus inactivating agent. For example, the following treatment system: In one chamber, the photosensitizer virus inactivating agent adsorption medium comprises styrene-containing organic polymer particles and a non-woven fabric or filter of photosensitizer physico-adsorbing fibers (for example, polyolefin fibers) located under the particles. material.
在本发明的病毒灭活方法的一项实施方案中, 在进行病毒灭活时所述光敏 剂病毒灭活剂的浓度为 0.55-3.0μιηΟ1 /1。 一个高的光敏剂病毒灭活剂浓度有利于 减少灭病毒时间、 或 /和应用有利于保持生物活性的灭病毒条件 (例如较低的温 度、 较低的光强度、 等等)。 实际上, 含病毒灭活剂吸附介质的固相介质的副作 用是限制光敏剂病毒灭活剂浓度的一个因素。 在一项实施方案中, 所述病毒灭 活在低于室温的温度下进行, 例如在 4-19.5°C之间、优选 15-19.5°C之间、更优 选 15-17°C之间迸行。 In one embodiment of the virus inactivation method of the invention, the concentration of the photosensitizer virus inactivating agent when the virus is inactivated is from 0.55 to 3.0 μηη Ο 1 /1. A high photosensitizer virus inactivating agent concentration is beneficial to reduce the time to virus elimination, or / and to apply virus-eliminating conditions that are beneficial to maintain biological activity (eg lower temperature) Degree, lower light intensity, etc.). In fact, the side effect of a solid phase medium containing a viral inactivating agent adsorption medium is one factor limiting the concentration of the photosensitizer virus inactivating agent. In one embodiment, the virus inactivation is carried out at a temperature below room temperature, for example between 4-19.5 ° C, preferably between 15 and 19.5 ° C, more preferably between 15 and 17 ° C. Row.
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包含所述有机溶 剂病毒灭活介质、 有机溶剂病毒灭活剂吸附介质、 或 /和光敏剂物理吸附纤维。 这是一种双重病毒灭活方法。  In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises the organic solvent virus inactivating medium, an organic solvent virus inactivating agent adsorption medium, or/and a photosensitizer physical adsorption fiber. This is a dual virus inactivation method.
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包括碘病毒灭活 介质, 所述碘病毒灭活介质至少含碘、 碘吸附介质和钝化剂。  In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises an iodine virus inactivating medium, the iodine virus inactivating medium comprising at least iodine, an iodine adsorption medium, and a passivating agent.
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包含固相病毒灭 活剂和其上固定的钝化剂。 在本发明的实施例中,固相病毒灭活剂的例子为正电 荷深层过滤滤材 (:例如 Cuno公司 Zetaplus VR滤板)。其中,固相病毒灭活剂为基 于正电荷作用的固相病毒灭活剂。 尽管由于现有产品的数量的限制, 本发明实 施例中仅举出这项实施方案的一个例子, 然而本专业技术人员应当知道, 这一 方法是容易推及其它固相病毒灭活剂上的。  In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises a solid phase virus inactivating agent and a passivating agent immobilized thereon. In an embodiment of the present invention, an example of a solid phase virus inactivating agent is a positive charge deep filtration filter (e.g., Cuno Corporation Zetaplus VR filter plate). Among them, the solid phase virus inactivating agent is a solid phase virus inactivating agent based on a positive charge action. Although an example of this embodiment is exemplified in the embodiments of the present invention due to the limitation of the number of existing products, those skilled in the art should know that this method is easy to push on other solid phase virus inactivating agents. .
在本发明的上述病毒灭活方法的一项实施方案中, 所述病毒灭活介质的高 度大于 2cm小于 6cm, 而所述病毒灭活剂吸附介质的高度大于 1cm小于 3cm。 为了安全, 使用过量的吸附介质是一种公知的做法, 然而过量的吸附介质又增 大副作用风险。  In an embodiment of the above virus inactivation method of the present invention, the virus inactivating medium has a height greater than 2 cm and less than 6 cm, and the virus inactivating agent adsorption medium has a height greater than 1 cm and less than 3 cm. For safety, it is a well-known practice to use an excess of the adsorption medium, however, excessive adsorption medium increases the risk of side effects.
在本发明的病毒灭活方法的一项实施方案中, 所述固相介质包括深层过滤 滤材,所述深层过滤滤材含所述病毒灭活剂吸附介质,且具有下述特征: A).平均密 度大于 0.25g/cm3; 和 B).灰分小于 1%。根椐本发明实施例,控制灰分对于副作用 最小化有着重要的意义。 滤材中除所述纤维外, 还可以含有胶粘剂、 增强剂 (例 如聚烯烃)、 等等。 较高密度的滤材可具有较高吸附能力。 在一项实施方案中, 所述固相介质包括下述一种滤材或它们的类似物: Seitz-BiolO、 Seitz-Bio20、 Seitz-bio40 、 Seitz-SupradurlOO、 Seitz-Supradur200 、 Seitz-Supradur500 、 Seitz-Supradurl000。在一项优选方案中, 所述滤材平均密度大于 In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises a depth filter medium, the deep filter medium containing the virus inactivating agent adsorption medium, and having the following characteristics: A) The average density is greater than 0.25 g/cm 3 ; and B). The ash content is less than 1%. According to an embodiment of the invention, controlling ash has important implications for minimizing side effects. The filter medium may contain, in addition to the fibers, an adhesive, a reinforcing agent (e.g., polyolefin), and the like. Higher density filter media can have higher adsorption capacity. In one embodiment, the solid phase medium comprises one of the following filters or their analogs: Seitz-BiolO, Seitz-Bio20, Seitz-bio40, Seitz-SupradurlOO, Seitz-Supradur200, Seitz-Supradur500, Seitz -Supradurl000. In a preferred embodiment, the average density of the filter media is greater than
Seitz-SupradurlO0、 Seitz-Supradur200)o 在本发明方法的一项实施方案中, 所述 深层过滤滤材的总高度大于 6mm。 Seitz-SupradurlO0, Seitz-Supradur 200) o In an embodiment of the method of the invention, the depth of the depth filter media is greater than 6 mm.
本发明中, 钝化吸附介质中可含一种或多种钝化剂、 一种或多种吸附介质、 等等。 上述钝化吸附介质中的吸附介质包括: 病毒灭活剂吸附介质、 病毒灭活 介质中的病毒灭活剂吸附介质、 和白细胞去除介质。 在本发明的一项实施方案 中, 所述病毒灭活剂吸附介质含下述一种或多种吸附剂: 活性碳、 无机吸附剂、 有机高分子化学吸附剂、 有机高分子亲和吸附剂、 有机髙分子物理吸附剂。 有 机高分子亲和吸附剂包括固定有配体 (例如: C6-C18碳链、 含肼的端基或病毒 RNA、或 DNA仿制品)的粒子或纤维。这些吸附剂对于病毒灭活剂有吸附功能, 但往往也具有较强的不需要的反应活性。 In the present invention, the passivating adsorption medium may contain one or more passivating agents, one or more adsorbing media, and the like. The adsorption medium in the passivation adsorption medium includes: a virus inactivating agent adsorption medium, a virus inactivating agent adsorption medium in the virus inactivating medium, and a leukocyte removal medium. In an embodiment of the present invention, the virus inactivating agent adsorption medium contains one or more of the following adsorbents: activated carbon, inorganic adsorbent, Organic polymer chemical adsorbent, organic polymer affinity adsorbent, organic germanium molecular physical adsorbent. The organic polymer affinity adsorbent includes particles or fibers to which a ligand (for example, a C6-C18 carbon chain, a hydrazine-containing end group or a viral RNA, or a DNA replica) is immobilized. These adsorbents have an adsorption function for virus inactivating agents, but tend to have strong undesired reactivity.
在本发明的病毒灭活方法的一项实施方案中, 所述钝化吸附介质中的吸附介 质包括活性炭。在本发明的实施例中,活性碳以不同的形式出现,例如活性碳粉、 活性碳毡、 含活性碳的滤材、 等等。  In an embodiment of the virus inactivation method of the present invention, the adsorbing medium in the passivating adsorption medium comprises activated carbon. In the embodiments of the present invention, activated carbon occurs in various forms such as activated carbon powder, activated carbon felt, activated carbon-containing filter material, and the like.
在本发明的病毒灭活方法的一项实施方案中, 所述钝化吸附介质中的吸附介 质包括离子交换吸附介质。离子交换吸附介质包括:硅藻土、珍珠岩、玻璃纤维、 等等无机吸附剂; 离子交换树脂、 离子交换纤维、 等等有机高分子化学吸附剂。  In an embodiment of the virus inactivation method of the present invention, the adsorbing medium in the passivating adsorption medium comprises an ion exchange adsorption medium. The ion exchange adsorption medium includes: an inorganic adsorbent such as diatomaceous earth, perlite, glass fiber, etc.; an organic polymer chemical adsorbent such as an ion exchange resin, an ion exchange fiber, or the like.
在本发明的病毒灭活方法的一项实施方案中, 所述钝化吸附介质中的吸附介 质包括上述物理吸附介质 (有机溶剂病毒灭活剂吸附介质、 光敏剂物理吸附纤 维)。 对物理吸附介质的钝化,有可能进一步减小副作用。  In an embodiment of the virus inactivation method of the present invention, the adsorption medium in the passivation adsorption medium comprises the above physical adsorption medium (organic solvent virus inactivating agent adsorption medium, photosensitizer physical adsorption fiber). Passivation of the physical adsorption medium makes it possible to further reduce side effects.
在本发明实施例中, 上述钝化剂优选可注射于人体的有机物。 这些钝化剂 的特征是: 1).本身是相对隋性的物质; 2).可被固定在固相介质上抑制其副作用 而对其功能没有有意义的不利影响; 3).即使有可控量的钝化剂从固相介质或其 它组成脱落至生物液体中, 其对生物液体的使用安全性不造成实质性的损害。  In the embodiment of the present invention, the passivating agent is preferably an organic substance that can be injected into a human body. These passivating agents are characterized by: 1) themselves being relatively inert; 2) can be immobilized on a solid phase medium to inhibit its side effects without meaningful adverse effects on its function; 3). The controlled amount of passivating agent is detached from the solid phase medium or other composition into the biological fluid, which does not cause substantial damage to the safety of use of the biological fluid.
在本发明的病毒灭活方法的一项实施方案中, 所述钝化吸附介质中的钝化 剂, 包括具有亲水基团或 /和亲油基团、 且可降低所述固相介质对所述生物液体 的副作用的有机物。 具有亲油基团的有机物水溶性差 (例如天然油旨、 有机溶剂 病毒灭活剂)、 而相当多具有亲水基团或具有亲水基团和亲油基团的有机物被认 为吸附力较差 (例如表面活性剂、羟基化合物、氨基酸), 不见用作固相介质的钝 化剂。 出人意料的是, 本发明的实施例中这类物质可用作本发明的钝化剂, 并 满足本发明的目的。  In an embodiment of the virus inactivation method of the present invention, the passivating agent in the passivation adsorption medium comprises a hydrophilic group or/and a lipophilic group, and the solid phase medium pair can be lowered An organic substance that describes the side effects of biological fluids. An organic substance having an oleophilic group is poorly water-soluble (for example, a natural oil, an organic solvent virus inactivating agent), and a considerable number of organic substances having a hydrophilic group or having a hydrophilic group and a lipophilic group are considered to have poor adsorption force ( For example, surfactants, hydroxy compounds, amino acids), no passivating agents are used as solid phase media. Surprisingly, such materials are useful as passivating agents in the practice of the present invention and are intended to serve the objectives of the present invention.
在本发明的一项实施方案中, 所述固相介质中的钝化剂含量大于 优选大于 1μιηο1/αη3。 对本发明的处理系统而言, 在某些情况中 只有当钝化剂的含量适当大时才能满足本发明的目的 (例如副作用最小化)。本发 明实施例中一些出乎意料的结果是, 当活性炭材料上的某些钝化剂的含量大于 5μηιο1/οιη3时, 其对病毒灭活剂 (例如亚甲蓝)的吸附仍然足够强。 至于钝化剂含 量的上限,则应根据其活性 (特异性吸附能力或病毒灭活能力)和所需的钝化程度 (例如对有无凝血因子的生物液体钝化程度可能要求不同)的不同,按公知方法加 以优化。 In an embodiment of the invention, the passivating agent content in the solid phase medium is greater than It is preferably greater than 1 μιηο1/αη 3 . For the treatment system of the present invention, the purpose of the present invention (e.g., minimizing side effects) can be met only in certain cases when the level of passivating agent is suitably large. Some unexpected results in the examples of the present invention are that the adsorption of viral inactivating agents (e.g., methylene blue) is still sufficiently strong when the amount of certain passivating agents on the activated carbon material is greater than 5 μηιο1/οιη 3 . As for the upper limit of the content of the passivating agent, it should be different according to its activity (specific adsorption capacity or virus inactivation ability) and the degree of passivation required (for example, the degree of passivation of biological fluids with or without clotting factors may be different) , optimized according to known methods.
在本发明的病毒灭活方法的一项实施方案中, 所述纯化剂包括天然油脂。 所 述天然油脂包括天然油脂和天然油脂衍生物。 这些物质均具有亲油基团。 在本 发明实施例中, 天然油脂包括植物油和磷脂, 其中: 植物油包括蓖麻油、 大豆 油、 茶油、 甘油; 磷脂包括脑磷脂、 卵磷脂和植物磷脂。 天然油脂衍生物的例 子包括植物油乳、 脂肪乳、 等等。 In an embodiment of the virus inactivating method of the present invention, the purifying agent comprises a natural fat. Place Natural oils and fats include natural oils and natural oils and fats. These materials all have an oleophilic group. In the embodiment of the present invention, the natural fats and oils include vegetable oils and phospholipids, wherein: vegetable oils include castor oil, soybean oil, tea oil, and glycerin; and phospholipids include cephalin, lecithin, and plant phospholipids. Examples of natural oil and fat derivatives include vegetable oil emulsions, fat emulsions, and the like.
在本发明的病毒灭活方法的一项实施方案中, 所述纯化剂包括羟基化合物。 羟基化合物具有亲水基团,其包括醇类 (例如、 乙醇、甘油、甘油氯化钠、等等)、 糖类、或 /和它们各自的衍生物。在一项实施方案中, 所述糖类包括单糖、 多糖、 或 /和聚多糖, 例如: 葡萄糖、 甘露醇(分子式 C6H1406)、 山梨醇、 甘油果糖、 蔗糖、 羟乙基淀粉、 香菇多糖、 等等。 In an embodiment of the virus inactivation method of the present invention, the purification agent comprises a hydroxy compound. The hydroxy compound has a hydrophilic group including alcohols (for example, ethanol, glycerin, glycerin sodium chloride, and the like), saccharides, or/and their respective derivatives. In one embodiment, the saccharide comprises a monosaccharide, a polysaccharide, or/and a polypolysaccharide, for example: glucose, mannitol (Molecular Formula C 6 H 14 0 6 ), Sorbitol, Glycerol Fructose, Sucrose, Hydroxyethyl Starch, lentinan, etc.
在本发明的病毒灭活方法的一项实施方案中, 所述纯化剂包括氨基酸。 氨基 酸具有亲水基团, 其包括胱氨酸、 赖氨酸、 酪氨酸、 甘氨酸、 精氨酸、 脯氨酸、 丝氨酸、 以及这些氨基酸的聚合物等等。 本发明实施例的令人吃惊的结果, 是 糖类物质或 /氨基酸和可被吸附在活性碳表面并降低活性碳对生物液体的副作 用。 聚氨基酸也具有类似作用。  In an embodiment of the virus inactivation method of the invention, the purification agent comprises an amino acid. The amino acid has a hydrophilic group including cystine, lysine, tyrosine, glycine, arginine, valine, serine, a polymer of these amino acids, and the like. The surprising result of embodiments of the present invention is the saccharide or /amino acid and can be adsorbed on the surface of the activated carbon and reduce the side effect of the activated carbon on the biological fluid. Polyamino acids also have a similar effect.
在本发明的病毒灭活方法的一项实施方案中, 所述纯化剂包括有机溶剂。 在 本发明实施例中,有机溶剂包括有机溶剂病毒灭活剂,例如:磷酸三丁酯 (TnBP)、 乙醚、 甘油、 碳酸乙酯、 乳酸乙酯、 苯甲酸苄酯、 等等。  In an embodiment of the virus inactivation method of the present invention, the purification agent comprises an organic solvent. In the embodiment of the present invention, the organic solvent includes an organic solvent virus inactivating agent such as tributyl phosphate (TnBP), diethyl ether, glycerin, ethyl carbonate, ethyl lactate, benzyl benzoate, and the like.
在本发明的一项实施方案中, 所述钝化剂包括多肽。 多肽具有亲水基团, 例如白蛋白、 血清、 和奶、 及它们各自的衍生物。  In an embodiment of the invention, the passivating agent comprises a polypeptide. The polypeptide has hydrophilic groups such as albumin, serum, and milk, and their respective derivatives.
在本发明的一项实施方案中, 所述钝化剂包括表面活性剂。 表面活性剂具 有亲水基团和亲油基团。 本发明发现, 大量表面活性剂、 尤其是分子结构中同 时具有亲水基团和亲油基团的表面活性剂, 可被吸附在活性碳表面并降低活性 碳对生物液体的副作用。亲水基团包括: -OH、 -OS03、 - (C¾CH20)3、 -N(CH3)2, -N(CH3)3、 -CH2COO、 -NH2; 亲油基团包括: 有机环基、 -(CH2 i-、 等等。 表面 活性剂的例子包括: Triton类表面活性物质、 吐温类表面活性物质、 胆酸钠、 四 氢糠醛聚乙二醇醚、 二甲基乙酰胺、 聚乙烯吡咯烷酮、 和二甲基亚砜。 部分表 面活性剂作为钝化剂的例子被给出在实施例中。 In an embodiment of the invention, the passivating agent comprises a surfactant. The surfactant has a hydrophilic group and a lipophilic group. The present inventors have found that a large number of surfactants, especially surfactants having both hydrophilic groups and lipophilic groups in the molecular structure, can be adsorbed on the surface of activated carbon and reduce the side effects of activated carbon on biological fluids. Hydrophilic groups include: -OH, -OS0 3 , - (C3⁄4CH 2 0) 3 , -N(CH 3 ) 2 , -N(CH 3 ) 3 , -CH 2 COO, -NH 2 ; lipophilic groups Including: organic ring groups, -(CH 2 i-, etc. Examples of surfactants include: Triton-based surface active substances, Tween-based surface active substances, sodium cholate, tetrahydrofurfural polyglycol ether, two Methylacetamide, polyvinylpyrrolidone, and dimethyl sulfoxide. Examples of partial surfactants as passivating agents are given in the examples.
在本发明的病毒灭活方法的一项实施方案中, 所述病毒灭活介质固定有有机 溶剂病毒灭活剂和有机溶剂钝化剂, 且所述有机溶剂钝化剂和有机溶剂病毒灭 活剂的总含量大于 0.2μπιο1/∞ι3、 优选大于 0.3μιηο1/αη3。 本发明的实施例说明, 只有当有机溶剂病毒灭活剂 (例如 ΤηΒΡ)含量大于某一程度, 有机溶剂才可既用 作病毒灭活剂进行有效的病毒灭活、 又可用作钝化剂使副作用最小化。 本发明 的实施例说明有机溶剂病毒灭活剂的含量大于 0.2 mol/cm3时可有此一特征。在 所述固相介质中, 还可含适量去污剂 D (例如含量大于 0.01 mol/cm3)或其它钝化 剂 (例如多糖)。 当 D含量足够大时 (例如大于 0.2Mmol/cm3), D也既可作病毒灭 活剂助剂、 又可作钝化剂。 In an embodiment of the virus inactivation method of the present invention, the virus inactivating medium is immobilized with an organic solvent virus inactivating agent and an organic solvent deactivator, and the organic solvent deactivator and the organic solvent virus are inactivated. The total content of the agent is greater than 0.2 μπιο1/∞ι 3 , preferably greater than 0.3 μιηο1/αη 3 . The examples of the present invention show that only when the content of the organic solvent virus inactivating agent (for example, ΤηΒΡ) is greater than a certain degree, the organic solvent can be used as a virus inactivating agent for effective virus inactivation and as a passivating agent. Minimize side effects. The embodiment of the present invention demonstrates that the content of the organic solvent virus inactivating agent is greater than 0.2 mol/cm 3 . In The solid phase medium may also contain an appropriate amount of detergent D (e.g., greater than 0.01 mol/cm 3 ) or other passivating agent (e.g., polysaccharide). When the D content is sufficiently large (for example, greater than 0.2 Mmol/cm 3 ), D can also be used as both a virus inactivating agent and a passivating agent.
在本发明的一项实施方案中, 所述钝化剂包括二种或二种以上上述钝化剂 的组合,例如:右旋糖酐 40/葡萄糖、右旋糖酐 40/白蛋白、白蛋白 /葡萄糖、 TnBP/ 吐温 -80/葡聚糖、 等等。  In an embodiment of the invention, the passivating agent comprises a combination of two or more of the above passivating agents, for example: dextran 40 / glucose, dextran 40 / albumin, albumin / glucose, TnBP / spit Wen-80/glucan, and so on.
在本发明的病毒灭活方法的一项实施方案中, 所述处理系统还含去白细胞 固相介质。 例如,本发明方法的一项实施方案, 其至少包括: A).提供单人份血液 组分; B).提供所述处理系统, 所述处理系统还含去白细胞固相介质; C).使所述 生物液体流过所述处理系统进行白细胞去除和病毒灭活或 /和病毒灭活剂吸附; D).从 C)中所述处理系统收集基本上不含所述病毒灭活剂和白细胞的单人份血 液组分。 在本发明方法的一项实施方案中, 所述去白细胞固相介质含所述有机 高分子物理吸附剂。  In one embodiment of the virus inactivation method of the invention, the treatment system further comprises a leukocyte-free solid phase medium. For example, an embodiment of the method of the present invention comprises at least: A) providing a single blood component; B) providing said treatment system, said treatment system further comprising a leukocyte solid phase medium; C). Flowing the biological fluid through the treatment system for leukocyte removal and virus inactivation or/and viral inactivating agent adsorption; D). The treatment system from C) is substantially free of the virus inactivating agent and A single blood component of white blood cells. In an embodiment of the method of the present invention, the leukocyte-removing solid phase medium comprises the organic polymeric physical adsorbent.
在本发明的病毒灭活方法的一项实施方案中, 所述去白细胞固相介质包括所 述病毒灭活剂吸附介质。  In an embodiment of the virus inactivation method of the present invention, the leukocyte-removing medium medium comprises the virus inactivating agent adsorption medium.
在本发明的病毒灭活方法的一项实施方案中, 所述生物液体包括单人份血 液组分。 在本发明的病毒灭活方法的一项实施方案中, 所述有机溶剂病毒灭活 介质中有机溶剂病毒灭活剂的含量,等于或小于 3 X (lOppmX所述单人份血液 组分的体积 ml)、或小于(lOppmX所述单人份血液组分的体积 ml)。实际上, 限 制有机溶剂病毒灭活剂的含量, 可能有助于免除最终产品中有机溶剂病毒灭活 剂的检测。  In one embodiment of the virus inactivation method of the invention, the biological fluid comprises a single human blood component. In an embodiment of the virus inactivation method of the present invention, the content of the organic solvent virus inactivating agent in the organic solvent virus inactivating medium is equal to or less than 3 X (10 ppm of the volume of the single blood component) Ml), or less than (lOppmX the volume of the single-part blood component ml). In fact, limiting the amount of organic solvent virus inactivating agent may help to eliminate the detection of organic solvent virus inactivating agents in the final product.
本发明的一种有效减小病毒危害的处理系统, 其为上述病毒灭活方法中所述 的处理系统。  A treatment system for effectively reducing virus hazards of the present invention is the treatment system described in the above virus inactivation method.
本发明的一种有效减小病毒危害的装置, 其至少含上述的处理系统。 所述装 置包括柱子、 滤器、 试剂盒、 等等。 在本发明实施例中, 一些所述装置还含病 毒灭活剂加入结构、 或 /和病毒灭活反应场所、 或 /和洗涤液。  An apparatus for effectively reducing virus hazards of the present invention, comprising at least the above-described processing system. The device includes a column, a filter, a kit, and the like. In an embodiment of the invention, some of the devices further comprise a viral inactivating agent addition structure, or/and a virus inactivation reaction site, or/and a wash solution.
在本发明的装置的一项实施方案中, 装置为单人份血液组分处理试剂盒。 实施例  In one embodiment of the device of the present invention, the device is a single human blood component treatment kit. Example
以下将参考实施例对本发明进行更为详细地描述。 但应认识到, 本发明实 施例仅给出本发明具体实施方式的个别情况的例子。 本专业的技术人员应当知 道,本发明不限于这些给定的实施模式(例如给定的过程、参数和组合)。因为, 本发明的内容是明确的, 但具体的过程、 参数和组合则可以是多变的。 以下实施例中, 所用试剂和材料均为市场有售的试剂和材料。 其中, 所用 吸附介质分别为: The invention will be described in more detail below with reference to examples. However, it should be understood that the embodiments of the invention are merely illustrative of the individual aspects of the embodiments of the invention. Those skilled in the art will appreciate that the present invention is not limited to these given modes of implementation (e.g., given processes, parameters, and combinations). Because the content of the invention is clear, the specific processes, parameters and combinations can be varied. In the following examples, the reagents and materials used were all commercially available reagents and materials. Among them, the adsorption media used are:
1) .活性碳: 所用含活性碳的病毒灭活剂吸附介质分别为活性碳粉、 活性碳 毡(ZC-1200A, 中国紫川炭纤维有限公司)和活性碳滤板 (AKS 5和 A S 6, 德 国 Seitz公司)。  1) Activated Carbon: The activated media for the activated carbon-containing virus inactivating agent are activated carbon powder, activated carbon felt (ZC-1200A, China Zichuan Carbon Fiber Co., Ltd.) and activated carbon filter plates (AKS 5 and AS 6, respectively). Seitz, Germany).
2) .离子交换吸附剂: 所用含离子交换吸附剂的病毒灭活剂吸附介质分别为 硅氧化物粒子和玻璃纤维。 所用含硅氧化物粒子的物质为含珍珠岩的滤板 (Zetaplus Delipid, Cuno公司)。  2). Ion exchange adsorbent: The virus inactivating agent adsorption medium containing the ion exchange adsorbent is silicon oxide particles and glass fibers, respectively. The material containing the silicon oxide particles was a perlite-containing filter plate (Zetaplus Delipid, Cuno Corporation).
3 亲和吸附剂: 所用含亲和吸附剂的病毒灭活剂吸附介质为 C18反相凝胶 (美国 Water公司)。  3 Affinity adsorbent: The virus inactivating agent adsorption medium containing the affinity adsorbent is C18 reverse phase gel (Water Company, USA).
4).物理吸附介质。 其中,所用物理吸附介质分别为:  4). Physical adsorption medium. Among them, the physical adsorption media used are:
(1) .光敏剂物理吸附纤维,包括天然纤维、纤维素基衍生纤维、非功能化合 成纤维。所用天然纤维包括植物纤维、 动物纤维 (例如蚕丝)。所用植物纤维包括 棉纤维 (例如无脂棉)、木纤维 (在滤板中,例如 Seitz Bio系列滤板)、纸浆 (例如纸)。 所用非功能化合成纤维包括亲和纤维和离子交换纤维以外的合成纤维, 例如聚 酯纤维、 聚丙烯纤维 (例如超细聚丙烯吸油纤维)、 聚酰氨纤维、 聚氨酯纤维(均 为无纺布)。尤其是聚烯烃纤维,其含于以下实施例所用作光敏剂物理吸附介质的 滤材中,例如 Seitz Supradur 80、 Seitz Supradur500 Seitz Eco 1000、 等等。  (1) Photosensitive agents Physically adsorbed fibers, including natural fibers, cellulose-based derived fibers, and non-functionalized synthetic fibers. The natural fibers used include plant fibers, animal fibers (e.g., silk). The plant fibers used include cotton fibers (e.g., non-fat cotton), wood fibers (in filter plates such as Seitz Bio series filter plates), and pulp (e.g., paper). The non-functionalized synthetic fibers used include synthetic fibers other than affinity fibers and ion exchange fibers, such as polyester fibers, polypropylene fibers (for example, ultrafine polypropylene oil absorbing fibers), polyamide fibers, and polyurethane fibers (both non-woven fabrics). ). In particular, polyolefin fibers are contained in the filter material used as the sensitizer physical adsorption medium in the following examples, such as Seitz Supradur 80, Seitz Supradur 500 Seitz Eco 1000, and the like.
(2) . 有机溶剂物理吸附介质,包括纤维 (有机溶剂物理吸附纤维)和大孔吸附 树脂 (有机溶剂物理吸附树脂)。所用纤维含有机溶剂病毒灭活剂的有机高分子物 理吸附剂,例如天然纤维 (例如 Seitz Bio 系列滤板)、 聚烯烃纤维 (:例如 Seitz Supradur 80, Seitz Supradur500, Seitz Eco 1000、 等等)、 等等。 所用大孔吸附 树脂 (有机溶剂物理吸附树脂)包括聚苯乙烯颗粒 (例如 Amberlite XAD- 7HP、 AmberliteXAD-16 ), 酚醛缩聚型颗粒、聚丙烯酸酯颗粒和含聚烷酯大孔吸附树 脂。 Amberlite XAD- 7HP为极性聚苯乙烯大孔吸附树脂颗粒, Amberlite XAD- 16 为非极性聚苯乙烯大孔吸附树脂颗粒。 所用颗粒具有下述一种或多种特征: A). 平均粒径 5-1000μιη; Β).比表面积 100-2000m 干粉; C).平均孔径小于 5-600A; D).排除体积小于 50000分子量。 部分所用多孔颗粒固相介质的球形蛋白质排阻 下限分子量小于 5000。 (2) Organic solvent physical adsorption medium, including fibers (organic solvent physical adsorption fibers) and macroporous adsorption resins (organic solvent physical adsorption resins). The fiber used contains an organic polymer physical adsorbent containing an organic solvent virus inactivating agent, such as natural fibers (for example, Seitz Bio series filter plates), polyolefin fibers (such as Seitz Supradur 80, Seitz Supradur 500, Seitz Eco 1000, etc.), and many more. The macroporous adsorption resin (organic solvent physical adsorption resin) used includes polystyrene particles (for example, Amberlite XAD-7HP, Amberlite XAD-16), phenolic polycondensation type particles, polyacrylate particles, and polyalkyl ester-containing macroporous adsorption resin. Amberlite XAD- 7HP is a polar polystyrene macroporous resin particle, and Amberlite XAD-16 is a non-polar polystyrene macroporous resin particle. The particles used have one or more of the following characteristics: A). Average particle size 5-1000 μιη ; Β). Specific surface area 100-2000 m dry powder; C). Average pore diameter less than 5-600 A; D) Excluding volume less than 50,000 molecular weight . The porous protein solid phase medium used in part has a spherical protein exclusion lower limit molecular weight of less than 5,000.
以下实施例中,上述光敏剂物理吸附纤维或有机溶剂物理吸附纤维包括纤 维丝、 无纺布、 布、 和深层过滤滤材。 所用含光敏剂物理吸附纤维或有机溶剂 物理吸附纤维的深层过滤滤材,平均密度大于 0.25g/cm3、 且灰分小于 1%, 包括 多种德国 Seitz公司产品 (KS80、 K900、 Supradur 80 > P30、 Eco 1000、 Permadur 0/400A、 T2600、 Bio20、 Bio40、 Bio60> Supradur 100、 等等)。 在本发明中, 前 缀 Seitz-是指德国 Sdtz公司产品。 专业人员应当知道, 其它公司的类似产品有 类似吸附性质。 部分所用深层过滤滤材固相介质的通透性小于 200L/m2min, 它 们分别为: Seitz-Supradur 100、 Seitz-biolO 、 Seitz-bio 400 In the following examples, the photosensitizer physical adsorbing fiber or the organic solvent physically adsorbing fiber comprises a fiber filament, a nonwoven fabric, a cloth, and a depth filter medium. The deep filter media containing the photosensitizer physically adsorbing fibers or organic solvent physically adsorbing fibers, the average density is greater than 0.25 g/cm 3 , and the ash content is less than 1%, including various German Seitz products (KS80, K900, Supradur 80 > P30) , Eco 1000, Permadur 0/400A, T2600, Bio20, Bio40, Bio60> Supradur 100, etc.). In the present invention, the prefix Seitz- refers to the product of the German company Sdtz. Professionals should be aware that similar products from other companies have similar adsorption properties. The permeability of some of the deep filter media used in the solid phase media is less than 200 L/m 2 min, which are: Seitz-Supradur 100, Seitz-biolO, Seitz-bio 40 0
下述实施例中, 所用钝化剂包括:  In the following examples, the passivating agent used includes:
1)具有亲油基团的天然油脂或 /和有机溶剂,例如: 蓖麻油、大豆油、茶油、 脂肪乳、 磷酸三丁脂 (本发明中简称 ΤπΒΡ)、 乙醚。  1) A natural fat or oil having an oleophilic group or/and an organic solvent, for example, castor oil, soybean oil, tea oil, fat milk, tributyl phosphate (abbreviated as ΤπΒΡ in the present invention), and diethyl ether.
2)具有亲水基团和亲油基团的表面活性剂, 例如: 胆酸钠、 Triton Χ100、 吐温 80、 和聚乙烯吡咯垸酮。  2) Surfactants having a hydrophilic group and an oleophilic group, for example: sodium cholate, Triton® 100, Tween 80, and polyvinylpyrrolidone.
3) . 具有亲水基团的烃基化合物, 例如: 甘油、葡萄糖、麦芽糖、葡聚糖 (右 旋糖酐)、 水溶性纤维素、 甘露醇(分子式 C6H1406)、 山梨醇、 羟乙基淀粉、 香 菇多糖。 3) Hydrocarbyl compounds having a hydrophilic group, such as: glycerin, glucose, maltose, dextran (dextran), water-soluble cellulose, mannitol (C 6 H 14 0 6 ), sorbitol, hydroxyethyl Starch, lentinan.
4) . 具有亲水基团的氨基酸, 例如: 脯氨酸、 精氨酸和甘氨酸。  4) . Amino acids having a hydrophilic group, such as: valine, arginine and glycine.
5) . 具有亲水基团的多肽, 例如: 人白蛋白 (北京天坛生物制品股分有限公 司)、 补血康 (德国 Biotest公司)和牛奶。  5). Polypeptides with hydrophilic groups, such as: human albumin (Beijing Tiantan Biological Products Co., Ltd.), Buxuekang (Germany Biotest) and milk.
6) .上述钝化剂的复合钝化剂, 例如: 右旋糖酐 40/葡萄糖、 右旋糖酐 40/白 蛋白、 白蛋白 /葡萄糖、 TnBP/吐温 -80/葡聚糖。 根据本发明下述实施例, 本专业 技术人员容易使用其它类似钝化剂来达到钝化目的。  6). A composite passivating agent for the above passivating agent, for example: dextran 40/glucose, dextran 40/albumin, albumin/glucose, TnBP/Tween-80/dextran. In accordance with the following examples of the invention, those skilled in the art will readily be able to use other similar passivating agents for the purpose of passivation.
下述实施例中, 所用病毒灭活剂包括:  In the following examples, the virus inactivating agent used includes:
1) .有机溶剂病毒灭活剂或有机溶剂病毒灭活剂 /去污剂,例如. ·磷酸三丁脂、 乙醚、 TnBP/吐温 -80、 TnBP/Triton X100、 TnBP/胆酸钠、 乙醚 /吐温 -80。  1). Organic solvent virus inactivating agent or organic solvent virus inactivating agent/detergent, for example. · Tributyl phosphate, diethyl ether, TnBP/Tween-80, TnBP/Triton X100, TnBP/sodium cholate, diethyl ether /Tween 80.
2) .光敏剂病毒灭活剂, 例如染料类病毒灭活剂或补骨脂素类病毒灭活剂。 所用染料类病毒灭活剂分别为亚甲蓝、 甲基紫、 甲苯胺蓝。 所用补骨脂素类病 毒灭活剂分别为补骨脂素和 8-methOXyprOTalen。专业人员应当知道,其它补骨脂 素与所用补骨脂素具有几乎相同的吸附性质。 2) Photosensitizer virus inactivating agents, such as dye virus inactivating agents or psoralen virus inactivating agents. The dye-like virus inactivating agents used were methylene blue, methyl violet, and toluidine blue, respectively. The psoralen virus inactivating agents used were psoralen and 8-m e th OX ypr OT al en , respectively . It should be understood by the skilled person that other psoralens have almost the same adsorption properties as the psoralen used.
3) .碘。 特别是碘 -PVPP滤板 (Sdtz公司)。  3) . Iodine. Especially iodine-PVPP filter plates (Sdtz).
4) . 固相病毒灭活剂, 例如 Zetaplus VR滤材 (Cuno公司:)。  4) . Solid phase virus inactivating agents, such as Zetaplus VR filter (Cuno:).
在以下的实施例中, 一般情况下病毒灭活的实施条件为: pH为 2.0-12.0; 温度为 -5至 6CTC ; 含病毒灭活剂的固相介质和生物液体的数量根据病毒灭活实 验来优选 (例如, 固相介质体积 /生物液体体积在 1%至 30%之间); 病毒灭活流体 动力学条件根据病毒灭活实验来优选(例如线性流速 0.1-lOcm/分钟、压力 0.1-5 公斤 /cm2); 含病毒灭活剂吸附介质的固相介质和生物液体的数量根据病毒灭活 剂去除实验来优选 (例如, 固相介质体积 /生物液体体积在 1%至 30%之间); 固- 液相吸附反应条件的确定, 基本上按照公知的规律进行优选, 例如: 生物液体 流过线速度越大, 吸附效率越小; 等等。 In the following examples, the conditions for virus inactivation are generally as follows: pH is 2.0-12.0; temperature is -5 to 6 CTC; amount of solid phase medium and biological liquid containing virus inactivating agent according to virus inactivation experiment Preferably (for example, solid medium volume / biological liquid volume between 1% and 30%); virus inactivation hydrodynamic conditions are preferred according to virus inactivation experiments (eg linear flow rate 0.1-lOcm/min, pressure 0.1-) 5 kg/cm 2 ); the amount of solid phase medium and biological liquid containing the virus inactivating agent adsorption medium is preferably according to the virus inactivating agent removal experiment (for example, the solid medium volume/bioliquid volume is between 1% and 30%) Between) The determination of the liquid phase adsorption reaction conditions is preferably carried out in accordance with a well-known rule, for example: the greater the flow rate of the biological liquid, the smaller the adsorption efficiency;
在以下的实施例中, 病毒灭活效率按照公知的方法用模式脂包膜病毒 In the following examples, the virus inactivation efficiency was carried out according to a known method using a pattern lipid enveloped virus.
VSV (初始滴度大于 104·8)和仙台病毒 (初始滴度大于 105·1)来研究。下述实施例中, 各种生物活性的检测均采用公知的方法进行。 本发明中, 白细胞去除率按公知 的方法测定和计算。 VSV (initial titer greater than 10 4 · 8 ) and Sendai virus (initial titer greater than 10 5 · 1 ) were studied. In the following examples, the detection of various biological activities was carried out by a known method. In the present invention, the leukocyte removal rate is measured and calculated by a known method.
下述实施例制备的装置, 包括可用于有效减小单人分血液组分的病原体危害 的装置。 其至少含本发明的处理系统, 其中的室为常规滤筒或层析柱。 下述实 施例制备的有效减小单人分血液组分的病原体危害的滤器, 一般而言, 内径为 The devices prepared in the following examples include devices that can be used to effectively reduce the pathogen hazard of a single human blood component. It comprises at least the treatment system of the invention, wherein the chamber is a conventional filter cartridge or chromatography column. The filter prepared by the following examples effective in reducing the pathogen hazard of a single human blood component, generally, the inner diameter is
3cm, 固相介质装填高度 3-5cm, 滤器出口端还有一层 0.4μηι滤膜, 以防介质碎 片漏出。 下述实施例制备的有效减小单人分血液组分的病原体危害的柱子, 一 般而言, 内径为 lcm, 固相介质装填高度 3-5cm, 柱的出口端还有一层 0.4μπι滤 膜, 以防介质碎片漏出。 3cm, the solid phase media loading height is 3-5cm, and there is a 0.4μηι filter at the outlet end of the filter to prevent the media debris from leaking out. The column prepared by the following examples is effective for reducing the pathogen hazard of a single blood component, generally having an inner diameter of 1 cm, a solid phase medium loading height of 3-5 cm, and a 0.4 μm filter at the outlet end of the column. In case the media debris leaks out.
实施例 1本发明的钝化固相介质的制备 Example 1 Preparation of passivated solid phase medium of the present invention
下述实施例给出若干较详细的例子。  The following examples give a few more detailed examples.
实施例 1.1含钝化剂的病毒灭活剂吸附介质 Example 1.1 Virus inactivating agent adsorption medium containing passivating agent
本实施例中制备的固相介质为吸附介质 /钝化剂复合物。  The solid phase medium prepared in this example is an adsorption medium/passivator composite.
1. 1.1 制备方法 1. 1.1 Preparation method
i).制备钝化剂分散处理系统  i). Preparation of passivation agent dispersion treatment system
本实施例中, 钝化剂以优选的浓度 (例如, 天然油脂: 0.2-0.5%; 有机溶剂: 0.3-2.0%; 表面活性剂: 1.0-3.0%; 多肽: 1.0-10.0%; 氨基酸: 3.0-10.0%; 等等) 分散在水溶液 (例如 PBS 缓冲液)中。 然而须知, 按照所用钝化剂的性质, 其它 介质 (例如有机溶剂病毒灭活剂:)也可作为钝化剂分散介质来制备钝化剂分散处 理系统 (例如溶液、悬浮液)。这时, 分散处理系统浓度 (w/v)可在 0.1%-50°/。之间。  In this embodiment, the passivating agent is at a preferred concentration (for example, natural fat: 0.2-0.5%; organic solvent: 0.3-2.0%; surfactant: 1.0-3.0%; polypeptide: 1.0-10.0%; amino acid: 3.0 -10.0%; etc.) Disperse in an aqueous solution (eg PBS buffer). However, it is to be noted that other media (e.g., organic solvent virus inactivating agents) may also be used as a passivating agent dispersion medium to prepare a passivator dispersion treatment system (e.g., solution, suspension) depending on the nature of the passivating agent used. At this time, the concentration of the dispersion treatment system (w/v) may be from 0.1% to 50°/. between.
低水溶性有机物钝化剂的分散, 可以采用公知技术。 天然油脂在水溶液中 的分散的方法很多, 例如乳化法、 除去空气法、 等等。 本实施例的一个方案, 还在分散介质中加入表面活性剂。 表面活性剂 (例如吐温 80、 Triton X 100)的使 用除有利于上述钝化剂、 特别是天然油脂或 /和有机溶剂病毒灭活剂在水中的分 散外, 有时还可作为洗脱剂来控制上述钝化剂在吸附介质上的吸附。  The dispersion of the low water-soluble organic substance deactivator can be carried out by a known technique. There are many methods for dispersing natural fats and oils in aqueous solutions, such as emulsification, air removal, and the like. In one embodiment of this embodiment, a surfactant is also added to the dispersion medium. The use of surfactants (eg Tween 80, Triton X 100) in addition to facilitating the dispersion of the above-mentioned passivating agents, in particular natural oils or/and organic solvent virus inactivating agents, in water, sometimes as an eluent The adsorption of the above passivating agent on the adsorption medium is controlled.
(2).将钝化剂固定至吸附介质上  (2) Fixing the passivating agent to the adsorption medium
本实施例中, 钝化剂在吸附介质上的固定是、 或基本上是通过吸附来进行 的。但需知道, 也可以选择其它固定方式 (例如共价键合)。 例如, 液态钝化剂也 可直接与吸附介质接触来进行吸附反应。 如果使用粉状吸附介质 (例如活性炭),则将其加入上述制备的钝化剂分散体 系中进行吸附反应。如果使用块状吸附介质 (例如活性炭滤板), 则以上述制备的 钝化剂分散体系为流动相、 以吸附介质为固定相进行流动吸附反应。 In this embodiment, the passivating agent is fixed on the adsorbent medium, or substantially by adsorption. However, it is important to know that other fixing methods (such as covalent bonding) can also be selected. For example, the liquid passivating agent can also be directly contacted with the adsorption medium to carry out the adsorption reaction. If a powdery adsorption medium (for example, activated carbon) is used, it is added to the above-prepared passivator dispersion system for adsorption reaction. If a bulk adsorption medium (for example, an activated carbon filter plate) is used, the passivation agent dispersion system prepared above is a mobile phase, and the adsorption medium is used as a stationary phase for a flow adsorption reaction.
所有的结合反应, 均是使用优化的钝化剂 /吸附介质比例、 在优化的反应条 件下迸行的。这些优化是按公知的结合技术 (例如吸附技术)迸行的。吸附反应的 条件包括: 反应物加入量、 pH、温度、 时间、某些添加剂 (例如表面活性剂、盐、 等等)的浓度、 流动相流速 (在流动吸附反应时)、 等等。 本专业的技术人员应当 知道通过控制这些条件来控制吸附反应, 从而获得所需的结果 (例如吸附量)。吸 附反应的均匀性也是一个需优先考虑的因素。  All binding reactions were carried out using optimized passivator/adsorption media ratios under optimized reaction conditions. These optimizations are performed in accordance with well-known bonding techniques (e.g., adsorption techniques). Conditions for the adsorption reaction include: reactant addition amount, pH, temperature, time, concentration of certain additives (e.g., surfactant, salt, etc.), mobile phase flow rate (at the time of flow adsorption reaction), and the like. Those skilled in the art will recognize that by controlling these conditions, the adsorption reaction is controlled to achieve the desired result (e.g., amount of adsorption). The uniformity of the adsorption reaction is also a priority.
本专业的专业人士应当知道,通过在固相载体 (例如层析胶)上固定病毒灭活 剂吸附剂 (例如 C18)、 然后再固定钝化剂也可以制备吸附介质 /钝化剂复合物。  Those skilled in the art will recognize that the adsorbent/passivator complex can also be prepared by immobilizing a viral inactivating adsorbent (e.g., C18) on a solid support (e.g., chromatography gel) followed by immobilization of the passivating agent.
(3).清洗未结合物  (3). Cleaning unconjugated
未固定、 或较弱吸附在吸附介质上的钝化剂和其它物质, 根据不同需要可 选用不同洗涤液 (例如 PBS缓冲液、 优选浓度的尿素溶液、 酒精溶液、 等等)进 行清洗或防脱性洗涤。  Passivation agents and other substances that are not fixed or weakly adsorbed on the adsorption medium, and may be washed or prevented by different washing liquids (for example, PBS buffer, preferred concentration of urea solution, alcohol solution, etc.) according to different needs. Sexual washing.
本实施例中制备的部份固相介质列于表 1中。  Some of the solid phase media prepared in this example are listed in Table 1.
上述制备的复合物、 特别是颗粒物质, 可单独用作固相介质 (例如表 1中的 A5-A30), 也可与其它组分 (例如增速物、 粘合剂、 增溶剂、 等等)混合后用作含 吸附介质和钝化剂的固相介质。例如表 1中, A1吸附介质 /钝化剂复合物和体积 百分比 10%的层析凝胶 (Sepharose FF, Pharmacia公司产品)混合; A2含吸附介 质 /钝化剂复合物和体积百分比 10%的珍珠岩混合。固相介质中的其它组分 (例如 增速物、 粘合剂、 增溶剂、 等等), 可以根据其所具有的不需要的反应活性, 按 照实际需要进行钝化 (例如, 形成其它固相组分 /钝化剂复合物), 或与吸附介质 混合后进行钝化 (例如, 形成固相介质 /钝化剂复合物:)。 它们的钝化方法与上述 吸附介质 /钝化剂复合物的制备方法中的钝化方法同。表 1中, A1是先形成固相 组分 /钝化剂复合物、 再混合形成固相介质制备的; A2是先混合形成固相介质, 再加入钝化剂来使不同组分钝化而制备的。  The composite prepared above, especially the particulate matter, can be used alone as a solid phase medium (for example, A5-A30 in Table 1), or with other components (such as a speed increasing substance, a binder, a solubilizing agent, etc.). After mixing, it is used as a solid phase medium containing an adsorption medium and a passivating agent. For example, in Table 1, A1 adsorption medium/passivator complex and 10% by volume chromatographic gel (Sepharose FF, product of Pharmacia) are mixed; A2 contains adsorption medium/passivator complex and 10% by volume Perlite mix. Other components in the solid phase medium (eg, speed-increasing substances, binders, solubilizers, etc.) can be passivated according to actual needs according to the undesired reactivity of the medium (for example, forming other solid phases) The component/passivator complex), or after mixing with the adsorption medium, is passivated (eg, forming a solid phase media/passivator complex:). Their passivation method is the same as that in the above-described method for preparing an adsorbent medium/passivator compound. In Table 1, A1 is prepared by first forming a solid phase component/passivator complex and then mixing to form a solid phase medium; A2 is first mixed to form a solid phase medium, and then a passivating agent is added to passivate the different components. Prepared.
表 1. 部分含钝化剂的病毒灭活剂吸附介质  Table 1. Part of virus inactivating agent adsorption medium containing passivating agent
复合物 吸附介质 钝化剂 复合物 吸附介质 钝化剂 Composite adsorption medium passivator composite adsorption medium passivation agent
A1 活性炭粉 蓖麻油 A21 AKS5 吐温 -80  A1 activated carbon powder castor oil A21 AKS5 Tween -80
A2 活性炭粉 右旋糖酐 A22 AKS5 聚乙烯吡咯烷酮  A2 activated carbon powder dextran A22 AKS5 polyvinylpyrrolidone
A3 ZC-1200A 人白蛋白 A23 AKS5 TnBP  A3 ZC-1200A Human Albumin A23 AKS5 TnBP
A4 AKS 6 脯氨酸 A24 AKS5 蓖麻油 A5 AKS 6 精氨酸 A25 A S5 大豆油 A4 AKS 6 Proline A24 AKS5 Castor Oil A5 AKS 6 arginine A25 A S5 soybean oil
A6 AKS 6 甘氨酸 A26 AKS5 脂肪乳  A6 AKS 6 glycine A26 AKS5 fat emulsion
A7 AKS 6 人白蛋白 A27 AKS5 TnBP/ Triton XI 00  A7 AKS 6 human albumin A27 AKS5 TnBP/ Triton XI 00
A8 AKS 6 葡聚糖 A28 AKS5 右旋糖酐 40/葡萄糖  A8 AKS 6 dextran A28 AKS5 dextran 40/glucose
A9 AKS 5 人白蛋白 A29 A S5 右旋糖酐 40/白蛋白  A9 AKS 5 human albumin A29 A S5 dextran 40/albumin
A10 AKS 5 补血康 A30 AKS5 TnBP/吐温 -80/葡聚糖  A10 AKS 5 Biankang A30 AKS5 TnBP/Tween-80/Glucan
All AKS 5 精氨酸 A31 Amberlite 右旋糖酐 40/甘氨酸  All AKS 5 arginine A31 Amberlite dextran 40/glycine
XAD- XAD-
7HP 7HP
A12 AKS 5 甘氨酸 A32 Amberlite 右旋糖酐 40/甘氨酸  A12 AKS 5 glycine A32 Amberlite dextran 40/glycine
XAD-1  XAD-1
A13 AKS 5 甘油 A33 玻璃纤维 人白蛋白  A13 AKS 5 Glycerin A33 Glass fiber Human albumin
A14 AKS 5 葡萄糖 A34 聚脂纤维 人白蛋白  A14 AKS 5 Glucose A34 Polyester Fiber Human Albumin
A15 AKS5 葡聚糖 A35 聚氨脂纤 人白蛋白  A15 AKS5 Dextran A35 Polyurethane Fiber Human Albumin
 Wei
A16 AKS5 麦芽、糖 A36 Zetaplus 人白蛋白  A16 AKS5 malt, sugar A36 Zetaplus human albumin
Delipid  Delipid
A17 AKS5 甘露醇 A37 Zetaplus 右旋糖酐 40/白蛋白  A17 AKS5 Mannitol A37 Zetaplus Dextran 40/Albumin
Delipid  Delipid
A18 AKS5 香菇多糖 A38 C18凝胶 人白蛋白  A18 AKS5 Lentinus Polysaccharide A38 C18 Gel Human Albumin
A19 AKS5 聚乙二醇 A39 C18凝胶 右旋糖酐 40/白蛋白  A19 AKS5 polyethylene glycol A39 C18 gel dextran 40/albumin
A20 AKS5 Triton A40 PVPP 滤 人白蛋白  A20 AKS5 Triton A40 PVPP Filtration Human Albumin
X100 板  X100 board
表 1 中,部分含钝化剂的病毒灭活剂吸附介质既是有机溶剂病毒灭活剂吸 附介质,又是光敏剂病毒灭活剂吸附介质 (例如含活性碳者和含大孔吸附树脂者)。  In Table 1, some of the virus inactivating agent adsorption media containing the passivating agent are both an organic solvent virus inactivating agent adsorption medium and a photosensitizer virus inactivating agent adsorption medium (for example, those containing activated carbon and macroporous adsorption resins). .
1.1.2鉴定  1.1.2 Identification
(1) . 固相介质中钝化剂含量的测定  (1) . Determination of the content of passivating agent in solid phase media
本实施例中,固相介质的钝化剂含量= (钝化剂加入总量 -未固定的钝化剂量 )/ 固相介质量。 其中, 未固定的钝化剂量通过公知的相关测定技术获得。 例如, 天然油脂的测定方法为析光仪法; 有机溶剂病毒灭活剂的测定方法为气相色谱 法; 表面活性剂的测定方法为气相色谱法; 多肽的测定方法为高压液相色谱法; 氨基酸的测定方法为高压液相色谱法; 等等。 本实施例制备的固相介质中, 大 多数情况下钝化剂的含量均大于 0.05μπιο1/αη3、 个别大于 0.4mmol/cm3In this embodiment, the passivating agent content of the solid phase medium = (total amount of passivating agent added - unfixed passivating dose) / amount of solid phase medium. Among them, the unfixed passivation dose is obtained by well-known correlation measurement techniques. For example, the method for measuring natural fats and oils is a spectrometer method; the method for measuring an organic solvent virus inactivating agent is gas chromatography; the method for measuring a surfactant is gas chromatography; and the method for determining a polypeptide is high pressure liquid chromatography; The method of determination is high pressure liquid chromatography; and the like. In the solid phase medium prepared in this example, in most cases, the content of the passivating agent is greater than 0.05 μπιο1/αη 3 and individually greater than 0.4 mmol/cm 3 .
(2) . 固相介质的特异性吸附的测定  (2) . Determination of specific adsorption of solid phase media
本发明中, 病毒灭活剂特异性吸附量 =(病毒灭活剂加入总量-未吸附的病毒 灭活剂量 y 固相介质体积。  In the present invention, the specific adsorption amount of the virus inactivating agent = (the total amount of the virus inactivating agent added - the unadsorbed virus inactivating dose y the volume of the solid phase medium).
本实施例中, 有机溶剂病毒灭活剂 (TnBP)和光敏剂病毒灭活剂 (亚甲蓝)分 别被用来测定特异性吸附。亚甲蓝的测定使用公知的分光光度计法。 TnBP的测 定使用公知的气相色谱法。 病毒灭活剂的吸附量, 按公知的动态吸附量测定方 法来测定。 测定所用装置为分别装有本实施例制备的固相介质和对照介质的柱 式容器 (体积 10ml)或滤器 (体积 10ml), 测定时 100ml病毒灭活剂溶液 (亚甲蓝浓 度 lOO g/ml; TnBP浓度 10mg /ml)以线速度为 0.3cm/分流过装置。 In this embodiment, the organic solvent virus inactivating agent (TnBP) and the photosensitizer virus inactivating agent (methylene blue) are divided into Do not be used to determine specific adsorption. The measurement of methylene blue uses a well-known spectrophotometer method. The measurement of TnBP uses a well-known gas chromatography. The amount of adsorption of the virus inactivating agent is measured by a known method for measuring the dynamic adsorption amount. The apparatus used for the measurement was a column container (volume 10 ml) or a filter (volume 10 ml) respectively containing the solid phase medium and the control medium prepared in the present example, and 100 ml of the virus inactivating agent solution (methylene blue concentration 100 g/ml) was measured. The TnBP concentration of 10 mg / ml was passed through the apparatus at a linear velocity of 0.3 cm / min.
表 1 中: 固相介质 A1-A35对光敏剂病毒灭活剂 (亚甲蓝)的吸附量均大于 O.Olmmol/cm3, 个别大于 0.02mmol/cm3、 个别甚至大于 O.lmmol/cm3 (例如 Al、 A2、 A32、 等等); 固相介质 A1-A30和 A36-A39的有机溶剂病毒灭活剂 (TnBP) 吸附量均大于 0.05mmol/cm3、 个别大于 O.lmmol/cm3、 个别甚至大于 0.3mmol/cm3; A1-A30对碘的吸附量大于 lmmol/cm3。此外, 部分上述固相介质 (特别是含活性碳者)对其它光敏剂病毒灭活剂、例如补骨脂素的特异性吸附, 亦 有类似结果。 In Table 1: The adsorption amount of the solid phase medium A1-A35 to the photosensitizer virus inactivating agent (methylene blue) is greater than O.Olmmol/cm 3 , and the individual is greater than 0.02 mmol/cm 3 , and sometimes even greater than 0.1 mmol/cm. 3 (for example, Al, A2, A32, etc.); the adsorption amount of the organic solvent virus inactivating agent (TnBP) of the solid phase media A1-A30 and A36-A39 is greater than 0.05mmol/cm 3 , and the individual is greater than O.lmmol/cm 3 , individual even greater than 0.3mmol / cm 3 ; A1-A30 adsorption of iodine greater than lmmol / cm 3 . In addition, some of the above solid phase media (especially those containing activated carbon) have similar results for the specific adsorption of other photosensitizer virus inactivating agents, such as psoralen.
表 1中, A33-A35还可作为去白细胞介质。  In Table 1, A33-A35 can also be used as a leukocyte-removing medium.
(3),固相介质的副作用的测定  (3) Determination of side effects of solid phase media
本实施例中, 副作用的测定包括测定非特异性吸附量或 /和 APTT (人血浆部 分凝血酶原活性)。 本实施例中, 非特异性吸附量= (指示试剂加入总量-未吸附的 指示试剂总量 )/固相介质体积。  In this example, the measurement of side effects includes measuring the amount of non-specific adsorption or / and APTT (human plasma fraction prothrombin activity). In this example, the amount of non-specific adsorption = (indicating the total amount of reagent added - the total amount of indicator reagent not adsorbed) / the volume of the solid phase medium.
本实施例中, 人白蛋白 (样品 C) (天坛生物制品股分有限公司)被用作非特 异性吸附指示试剂; 人血浆 (样品 D)的部分凝血酶原活性时间 (本发明中简称 APTT)被用作凝血处理系统变化的指示参数。 ΑΡΎΤ试剂盒购自中国医学科学院 成都输血研究所。  In this example, human albumin (sample C) (Tiantan Biological Products Co., Ltd.) was used as a non-specific adsorption indicating reagent; partial prothrombin activity time of human plasma (sample D) (abbreviated as APTT in the present invention) Used as an indicator of changes in the coagulation system. The sputum kit was purchased from the Chengdu Institute of Blood Transfusion, Chinese Academy of Medical Sciences.
与上述特异性吸附的测定方法相同, 副作用的测定也按公知的动态反应测 定方法进行。 测定时人白蛋白 (浓度 5%)或人血浆 (蛋白浓度 5.5%)与固相介质的 体积比在 3 : 1至 5: 1之间。  As in the above-described measurement method of specific adsorption, the measurement of side effects is also carried out by a known dynamic reaction measurement method. The ratio of human albumin (5%) or human plasma (5.5% protein) to the solid medium was between 3:1 and 5:1.
本实施例中制备的吸附介质 /钝化剂复合物与对照吸附介质比较, 副作用明 显下降,体现为: A).白蛋白吸附量 (mg/cm3)下降 25%以上,个别下降 50%以上 (:例 如白蛋白 /吸附介质复合物、植物油 /吸附介质复合物、糖 /吸附介质复合物、等等:); B).人血浆 APTT升高值下降 30%以上, 个别下降 100%以上 (例如白蛋白 /吸附介 质复合物、 植物油 /吸附介质复合物、 糖 /吸附介质复合物、 等等)。 Compared with the control adsorption medium, the adsorption medium/passivator composite prepared in this example has a significant decrease in side effects, which is reflected as: A). The albumin adsorption amount (mg/cm 3 ) decreases by more than 25%, and the individual decreases by more than 50%. (: for example, albumin/adsorption medium complex, vegetable oil/adsorption medium complex, sugar/adsorption medium complex, etc.); B). The increase in human plasma APTT is more than 30%, and the individual drop is more than 100% ( For example, albumin/adsorption medium complex, vegetable oil/adsorption medium complex, sugar/adsorption medium complex, etc.).
实施例 1.2含钝化剂的病毒灭活介质 (1) Example 1.2 Virus inactivating medium containing passivating agent (1)
本实施例中制备的固相介质为有机溶剂病毒灭活剂 /吸附介质 /钝化剂复合 物。  The solid phase medium prepared in this example is an organic solvent virus inactivating agent/adsorption medium/passivator compound.
1.2.1 制备方法 (1) .从吸附介质制备病毒灭活剂 /吸附介质复合物 1.2.1 Preparation method (1) Preparation of virus inactivating agent/adsorption medium complex from adsorption medium
(A).制备病毒灭活剂溶液或悬浮液  (A). Preparation of a virus inactivating agent solution or suspension
本实施例中, 以 PBS缓冲液为分散相, 按公知技术制备病毒灭活剂溶液或 悬浮液, 例如: TnBP/Triton X 100/水溶液 (5%TnBP, 5%Triton X 100); TnBP/吐 温 80/水溶液 (3% TnBP, 10%吐温 80浓度); β -丙内酯 /吐温 80/水溶液 (0.5% β - 丙内酯); 等等。  In this embodiment, a virus inactivating agent solution or suspension is prepared according to a known technique using PBS buffer as a dispersed phase, for example: TnBP/Triton X 100/water solution (5% TnBP, 5% Triton X 100); TnBP/spray Warm 80/water solution (3% TnBP, 10% Tween 80 concentration); β-propiolactone/Tween 80/water solution (0.5% β-propiolactone);
(Β).将病毒灭活剂固定至吸附介质上  (Β). Fix the virus inactivating agent to the adsorption medium
通过常规的固 -液相吸附反应条件进行, 将液相中的病毒灭活剂吸附在固相 吸附介质上。  The virus inactivating agent in the liquid phase is adsorbed on the solid phase adsorption medium by a conventional solid-liquid phase adsorption reaction condition.
(C).将钝化剂固定至吸附介质上  (C). Fixing the passivating agent to the adsorption medium
固定方法与实施例 1.1中 "将钝化剂固定至吸附介质上" 的方法相同。  The fixing method is the same as the method of "fixing the passivating agent to the adsorption medium" in the embodiment 1.1.
(2) . 从病毒灭活剂 /吸附介质复合物制备病毒灭活剂 /吸附介质 /钝化物复合 物  (2) . Preparation of virus inactivating agent / adsorption medium / passivation complex from virus inactivating agent / adsorption medium complex
本实施例中病毒灭活剂 /吸附介质复合物的制备方法, 与本实施例 (1)中 "将 病毒灭活剂固定至吸附介质上"的方法相同 (例如碘 -PVPP滤板)。 而将钝化剂固 定至病毒灭活剂 /吸附介质复合物上的方法, 则与实施例 1.1 中 "将钝化剂固定 至吸附介质上" 的方法相同。  The preparation method of the virus inactivating agent/adsorption medium complex in this embodiment is the same as the method of "fixing the virus inactivating agent to the adsorption medium" in the present embodiment (1) (e.g., iodine-PVPP filter plate). The method of fixing the passivating agent to the virus inactivating agent/adsorption medium composite is the same as the method of "fixing the passivating agent to the adsorption medium" in the embodiment 1.1.
本实施例制备的部分固相介质列于表 2中。 本实施例中的有机溶剂病毒灭 活剂 /吸附介质 /钝化剂复合物 (例如表 2中的 B1-B3)可单独用作、 或与其它组分 (例如增速物、粘合剂、 增溶剂、 等等)混合后用作含病毒灭活剂的固相介质。 其 钝化方法, 也可先混合形成固相介质再加入钝化剂钝化、 或钝化不同组分再使 它们混合。  Some of the solid phase media prepared in this example are listed in Table 2. The organic solvent virus inactivating agent/adsorption medium/passivator complex (for example, B1-B3 in Table 2) in this embodiment may be used alone or in combination with other components (for example, a speed increasing substance, a binder, The solubilizer, etc.) is used as a solid phase medium containing the virus inactivating agent after mixing. The passivation method may also be first mixed to form a solid phase medium, followed by passivation to passivate, or passivate the different components and then mix them.
1.2.2鉴定  1.2.2 Identification
本实施例制备的病毒灭活介质, 其病毒灭活效率的测定按上述方法进 ^1, 测定在室温进行, 线性流速小于 lml/cm/分, 生物液体与固相载体的体积比小于 100。 本实施例制备的病毒灭活剂 /吸附介质 /钝化剂复合物具有有效的病毒灭活 功能。 Virus inactivation medium prepared in the present embodiment, the measured efficiency of inactivated virus into which ^ 1 described above, measurement was carried out at room temperature, less than the linear flow rate of lml / cm / min, the volume of biological fluid with a solid support is less than 100. The virus inactivating agent/adsorption medium/passivator complex prepared in this example has an effective virus inactivating function.
本实施例制备的病毒灭活介质, 其钝化剂含量、 病毒灭活剂含量、 固相介 质副作用的测定方法与实施例 1中的相应方法相同。  The virus inactivating medium prepared in this example has the same method of determining the passivating agent content, the virus inactivating agent content, and the solid phase medium side effect as in the corresponding method in the first embodiment.
表 2中本实施例制备的病毒灭活介质,钝化剂的含量大于 0.05 mol/cm3、个 别大于 0.4mmol /cm3。 与对照病毒灭活剂 /吸附介质复合物比较, 病毒灭活效率 未因钝化而明显改变, 而副作用明显下降: A).白蛋白吸附量 (mg/cm3)下降 15% 以上,个别下降 30%以上; B).人血浆 APTT升高值下降 20%以上,个别下降 50% 以上。 The virus inactivating medium prepared in this example in Table 2 has a passivating agent content of more than 0.05 mol/cm 3 and an individual content of more than 0.4 mmol / cm 3 . Compared with the control virus inactivating agent/adsorption medium complex, the virus inactivation efficiency was not significantly changed by passivation, but the side effects were significantly decreased: A). The albumin adsorption amount (mg/cm 3 ) decreased by more than 15%, and decreased individually. More than 30%; B). The increase in human plasma APTT is more than 20%, and the individual is reduced by 50%. the above.
特别要指出的是, 在表 2的 B7或 B8中: A).所述病毒灭活剂包括有机溶剂 病毒灭活剂; B).所述钝化剂包括有机溶剂病毒灭活剂; 和 C).所述固相介质中 有机溶剂病毒灭活剂的含量大于 0.1mmol /cm3、甚至大于 0.3mmol/cm3 (例如 0.40 mmol/cm3)0 这时, 有机溶剂病毒灭活剂可既用作病毒灭活剂进行有效的病毒灭 活、 又可用作病毒灭活剂吸附介质 (例如活性碳)的钝化剂去使副作用最小化。 实施例 1.3含钝化剂的病毒灭活介质 (2) In particular, in B7 or B8 of Table 2: A). The virus inactivating agent comprises an organic solvent virus inactivating agent; B). the passivating agent comprises an organic solvent virus inactivating agent; and C The content of the organic solvent virus inactivating agent in the solid phase medium is greater than 0.1 mmol / cm 3 , or even greater than 0.3 mmol / cm 3 (for example, 0.40 mmol / cm 3 ) 0 at this time, the organic solvent virus inactivating agent can It can be used as a virus inactivating agent for effective virus inactivation, and can also be used as a passivating agent for a virus inactivating agent adsorption medium (for example, activated carbon) to minimize side effects. Example 1.3 Virus inactivating medium containing passivating agent (2)
本实施例制备的是固相病毒灭活剂 /钝化剂复合物。 所用固相病毒灭活剂分 别为 Cuno-Zetaplus VR灭病毒滤板和 Seitz-碘 -PVPP滤板。将钝化剂固定至固相 病毒灭活剂的方法,与实施例 U中 "将钝化剂固定至吸附介质上" 的方法相同。 制备物列于表 2中。  In this example, a solid phase virus inactivating agent/passivator complex was prepared. The solid phase virus inactivating agents used were Cuno-Zetaplus VR virus-killing filter plates and Seitz-iodine-PVPP filter plates. The method of fixing the passivating agent to the solid phase virus inactivating agent is the same as the method of "fixing the passivating agent to the adsorption medium" in Example U. The preparations are listed in Table 2.
本实施例制备的病毒灭活介质, 其病毒灭活效率、 钝化剂含量、 和副作用 的测定方法分别与实施例相应测定方法相同。  The virus inactivating medium prepared in this example has the same method for determining the virus inactivation efficiency, the depressant content, and the side effects, respectively, in accordance with the corresponding measurement methods of the examples.
表 2中本实施例制备的固相病毒灭活剂 /钝化剂复合物, 钝化剂的含量大于 0.05μπιο1/ ιι3。与对照固相病毒灭活剂比较,病毒灭活效率未因钝化而明显改变, 而副作用明显下降: Α).白蛋白吸附量 (mg/cm3)下降 5%以上; B).人血浆 APTT 升高值下降 10%以上。 The solid phase virus inactivating agent/passivator complex prepared in this example in Table 2 has a passivating agent content of more than 0.05 μπιο1/ιι 3 . Compared with the control solid phase virus inactivating agent, the virus inactivation efficiency was not significantly changed by passivation, but the side effects were significantly decreased: Α). The albumin adsorption amount (mg/cm 3 ) decreased by more than 5%; B). Human plasma The APTT rise is down more than 10%.
表 2.部分含病毒灭活剂的固相介质  Table 2. Part of solid phase media containing virus inactivating agent
复合物 吸附介质 病毒灭活剂 钝 化 剂  Complex adsorption medium virus inactivating agent passivator
B1 活性碳粉 S/D 精氨酸  B1 activated carbon powder S/D arginine
B2 活性碳粉 S/D 脯氨酸  B2 activated carbon powder S/D proline
B3 活性碳粉 S/D 甘氨酸  B3 activated carbon powder S/D glycine
B4 AKS5 S/D 人白蛋白  B4 AKS5 S/D Human Albumin
B5 A S5 S/D 牛奶  B5 A S5 S/D milk
B6 AKS5 S/D 补血康  B6 AKS5 S/D Bianxikang
B7 AKS5 S/D TnBP  B7 AKS5 S/D TnBP
B8 AKS5 S/D TnBP/ Triton XI 00  B8 AKS5 S/D TnBP/ Triton XI 00
B9 AKS5 S/D Triton XI 00  B9 AKS5 S/D Triton XI 00
B10 AKS5 S/D 葡萄糖  B10 AKS5 S/D Glucose
B11 AKS5 S/D 葡聚糖  B11 AKS5 S/D dextran
B12 PS吸附树脂 S D 右旋糖酐 40/葡萄糖  B12 PS adsorption resin S D dextran 40/glucose
B13* PVPP 碘 人白蛋白  B13* PVPP iodine human albumin
B14 PS**吸附树脂 S/D 右旋糖酐 40/葡萄糖  B14 PS** adsorption resin S/D dextran 40/glucose
B15 Zetaplus VR滤材 人白蛋白 B16 Zetaplus VR滤材 甘氨酸 B15 Zetaplus VR filter human albumin B16 Zetaplus VR filter material glycine
B13*: SEITS PVPP碘滤板; PS**: 聚苯乙烯 B13* : SEITS PVPP iodine filter plate; PS**: polystyrene
实施例 1. 4含钝化剂的病毒灭活剂吸附介质 /去白细胞固相介质 Example 1. 4 virus inactivating agent adsorption medium containing deactivating agent / leukocyte solid phase medium
本实施例中, 所用去白细胞介质-病毒灭活剂吸附介质分别为: 玻璃纤维、 棉纤维、聚脂纤维、聚氨基甲酸乙酯(polyurethane)无纺布、 Seitz-Supradur 100 聚丙烯纤维滤板; 所用钝化剂分别为上述钝化剂 (例如, 甘氨酸、 右旋糖酐 40/ 葡萄糖、 和它们的混合物)。  In this embodiment, the leukocyte-removing medium-virus inactivating agent adsorption medium used is: glass fiber, cotton fiber, polyester fiber, polyurethane nonwoven fabric, Seitz-Supradur 100 polypropylene fiber filter plate The passivating agent used is the above passivating agent (for example, glycine, dextran 40/glucose, and mixtures thereof).
本实施例制备的钝化剂 /病毒灭活剂吸附介质-去白细胞固相介质复合物,其 制备方法与实施例 1.1 中吸附介质 /钝化剂复合物的制备方法相同, 吸附反应在 去白细胞介质-病毒灭活剂吸附介质与钝化剂之间进行, 其中钝化剂含量和固相 介质副作用的测定方法, 分别与实施例 1.1中的相应测定的方法相同。  The passivating agent/viral inactivating agent adsorption medium-de-whitening cell solid phase medium composite prepared in this embodiment is prepared in the same manner as the adsorption medium/passivator complex in the embodiment 1.1, and the adsorption reaction is in the leukocyte-removing cell. The medium-viral inactivating agent is adsorbed between the adsorption medium and the passivating agent, and the method for determining the content of the passivating agent and the side effect of the solid phase medium is the same as that of the corresponding method in Example 1.1, respectively.
本实施例的制备物,其白细胞去除率可达 99%以上,亚甲兰去除率可达 95% 以上, 而其副作用比未结合钝化剂的相应去白细胞介质明显降低: A).白蛋白吸 附量 (mg/cm3)下降 15%以上; B).人血浆 APTT升高值下降 20%以上。 The preparation of the present embodiment has a leukocyte removal rate of over 99%, and a methylene blue removal rate of over 95%, and the side effects thereof are significantly lower than the corresponding leukocyte-removing medium without a passivating agent: A). Albumin The amount of adsorption (mg/cm 3 ) decreased by more than 15%; B). The increase in human plasma APTT decreased by more than 20%.
实施例 2.本发明的装置的制备 (1) Example 2. Preparation of the apparatus of the present invention (1)
本实施例制备的装置, 所用固相介质选自实施例 1、 特别是其中的实施例 1.1-1.4制备的钝化固相介质。  The apparatus prepared in this example, the solid phase medium used is selected from the passivated solid phase medium prepared in Example 1, especially the examples 1.1-1.4 therein.
本实施例的装置通过两种方法获得: A).将己制好的钝化固相介质装入室中; B).将固相介质装入室中再用钝化剂对其钝化, 钝化方法与实施例 1中 "将钝化 剂固定至吸附介质上"的方法相同。  The apparatus of this embodiment is obtained by two methods: A) loading a prepared passivated solid phase medium into the chamber; B) loading the solid phase medium into the chamber and then passivating it with a passivating agent, The passivation method is the same as the method of "fixing the passivating agent to the adsorption medium" in Example 1.
本实施例制备的装置, 其鉴定方法与实施例 1 中钝化固相介质的相应鉴定 方法相同。 一般而言, 其功能 (例如, 特异性吸附能力、 白细胞去除率、 病毒灭 活效率)和副作用分别与其所含钝化固相介质的功能和副作用一致。 即功能未因 加入钝化剂而有意义地降低, 而其副作用则明显下降。  The apparatus prepared in this embodiment has the same identification method as the corresponding identification method of the passivated solid phase medium in Embodiment 1. In general, its functions (e.g., specific adsorption capacity, leukocyte removal rate, virus inactivation efficiency) and side effects are consistent with the function and side effects of the passivated solid phase medium contained therein, respectively. That is, the function is not significantly reduced by the addition of a passivating agent, and its side effects are significantly reduced.
下述实施例给出若干较详细的例子。  The following examples give a few more detailed examples.
实施例 2.1病毒灭活剂除去装置 Example 2.1 Virus inactivating agent removal device
实施例 2.1.1 病毒灭活剂去除滤器 Example 2.1.1 Virus inactivating agent removal filter
本实施例中, 滤器中的固相介质选自钝化剂 /病毒灭活剂吸附介质 (纤维或 / 和滤材)复合物。例如, 钝化剂 /聚苯乙烯纤维复合物、钝化剂 /聚丙乙烯纤维复合 物、 钝化剂 /活性碳滤板复合物、 等等。  In this embodiment, the solid phase medium in the filter is selected from the group consisting of a passivating agent/viral inactivating agent adsorption medium (fiber or/and filter medium). For example, passivating agent/polystyrene fiber composite, passivating agent/polypropylene fiber composite, passivating agent/activated carbon filter plate composite, and the like.
实施例 2.1.2病毒灭活剂去除柱 Example 2.1.2 Virus inactivating agent removal column
本实施例中, 病毒灭活剂去除柱中的固相介质选自钝化剂 /病毒灭活剂吸附 介质 (颗粒)复合物。 例如, 钝化剂 /聚苯乙烯大孔吸附树脂复合物、 钝化剂 /活性 碳粉复合物、 等等。 In this embodiment, the solid phase medium in the virus inactivating agent removal column is selected from the group consisting of a passivating agent/viral inactivating agent. Medium (particle) composite. For example, passivating agent/polystyrene macroporous adsorption resin composite, passivating agent/activated carbon powder composite, and the like.
实施例 2.2病毒灭活装置 Example 2.2 Virus inactivation device
实施例 2.2.1病毒灭活滤器 Example 2.2.1 Virus inactivation filter
本实施例中, 病毒灭活滤器中的固相介质分别为: 1).固定有钝化剂的病毒 灭活介质 (:例如高含量 TnBP/土温 80/活性碳滤板复合物); 2). 固定有钝化剂的病 毒灭活介质和固定有钝化剂的病毒灭活剂吸附介质 (例如高含量 TnBP/土温 80/ 活性碳滤板复合物和糖-氨基酸纯化剂 /活性碳滤板复合物);以及 3).病毒灭活介 质和固定有钝化剂的病毒灭活剂吸附介质 (:例如 TnBP/土温 80/活性碳滤板复合 物和糖-氨基酸纯化剂 /活性碳滤板复合物:)。  In this embodiment, the solid phase medium in the virus inactivating filter is: 1) a virus inactivating medium immobilized with a passivating agent (for example, a high content of TnBP/earth temperature 80/activated carbon filter plate composite); a virus inactivating medium immobilized with a passivating agent and a virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, a high content of TnBP/earth temperature 80/activated carbon filter plate complex and a sugar-amino acid purifying agent/activated carbon) a filter plate complex; and 3) a virus inactivating medium and a virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, TnBP/earth temperature 80/activated carbon filter plate complex and sugar-amino acid purifying agent/activity Carbon filter plate composite :).
实施例 2.2.2病毒灭活柱 Example 2.2.2 Virus inactivation column
本实施例中, 病毒灭活柱中的固相介质分别为: 1).固定有钝化剂的病毒灭 活介质 (例如高含量 TnBP/土温 80/含苯乙烯的聚合物吸附树脂复合物); 2). 固定 有钝化剂的病毒灭活介质和固定有或无钝化剂的病毒灭活剂吸附介质 (例如高含 量 TnBP/土温 80/含苯乙烯的聚合物吸附树脂复合物和糖-氨基酸纯化剂 /活性碳 滤板复合物, 或高含量 TnBP/土温 80/含苯乙烯的聚合物吸附树脂复合物和含苯 乙烯的聚合物吸附树脂:); 以及 3).病毒灭活介质和固定有钝化剂的病毒灭活剂 吸附介质 (例如含苯乙烯的聚合物吸附树脂 /TnBP/土温 80复合物和钝化剂 /含苯 乙烯的聚合物吸附树脂复合物)。  In this embodiment, the solid phase medium in the virus inactivation column is: 1) a virus inactivating medium immobilized with a passivating agent (for example, a high content of TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite) 2). Virus inactivation medium with passivator immobilized and virus inactivating agent adsorption medium with or without passivating agent (eg high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin complex) And sugar-amino acid purifier/activated carbon filter plate composite, or high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite and styrene-containing polymer adsorption resin :); and 3). virus Inactivation medium and virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, styrene-containing polymer adsorption resin/TnBP/earth temperature 80 composite and passivating agent/styrene-containing polymer adsorption resin composite) .
实施例 2.3.去白细胞 /病毒灭活剂去除滤器 Example 2.3. Leukocyte-removing/viral inactivating agent removal filter
本实施例中, 去白细胞滤器中的固相介质选自钝化剂 /去白细胞介质 /病毒灭 活剂吸附介质复合物。 例如, 在滤筒中装入一层 T 2600滤板、 4 层钝化剂 /Seitz-Supradur 100滤板复合物, 出口端有一层 0.4μηι滤膜, 然后用 8%的右旋 糖酐循环泵入钝化 1 小时, 烘干消毒后备用。 该滤器可用于去白细胞和病毒灭 活剂 (例如亚甲蓝)去除。  In this embodiment, the solid phase medium in the leukocyte-removing filter is selected from the group consisting of a passivating agent / a leukocyte-removing medium / a virus inactivating agent adsorption medium complex. For example, a filter cartridge is packed with a T 2600 filter plate, 4 layers of passivator/Seitz-Supradur 100 filter plate composite, a 0.4 μηι filter at the outlet end, and then pumped into passivation with 8% dextran. Hours, dry and disinfected for use. This filter can be used to remove leukocytes and viral inactivating agents such as methylene blue.
实施例 3.本发明的装置的制备 (2) Example 3. Preparation of the device of the invention ( 2 )
本实施例制备的装置, 其中的固相介质含上述光敏剂物理吸附纤维,例如天 然纤维、聚酰氨纤维、聚氨酯纤维和滤板 (例如上述 Supmdur 100、 Supradur 500 Eco l000、 Permadur 0/400A T2600、 Bio20、 Bio40、 Bio60、 等等)。  The apparatus prepared in the embodiment, wherein the solid phase medium comprises the above-mentioned photosensitizer physical adsorption fiber, such as natural fiber, polyamide fiber, polyurethane fiber and filter plate (for example, Supmdur 100, Supradur 500 Eco l000, Permadur 0/400A T2600 described above). , Bio20, Bio40, Bio60, etc.).
本实施例制备的装置, 其鉴定方法与实施例 2制备的相应装置的鉴定方法 相同。 一般而言, 与现有的装置 (例如与含活性炭或无机吸附介质的装置)相比, 本实施例制备的装置有类似的光敏剂病毒灭活剂除去能力, 而有明显较低的副 作用 (例如, 蛋白吸附低 15%以上或 /和 APTT值升高值低 15%以上), 或有明显 较低的颗粒污染 (例如, 流过处理系统的生物液体中在显微镜下可见的直径大于The apparatus prepared in this embodiment has the same identification method as the corresponding apparatus prepared in the second embodiment. In general, the apparatus prepared in this example has a similar photosensitizer virus inactivating agent removal ability and has significantly lower side effects than the existing apparatus (for example, a device containing activated carbon or an inorganic adsorption medium). For example, protein adsorption is 15% lower or / and APTT value is lower than 15%), or there is obvious Lower particle contamination (for example, the diameter of the biological fluid flowing through the treatment system visible under the microscope is greater than
Ιμιη的颗粒数目低 15%以上)。 Ιμιη has a lower particle count of 15% or more).
下述实施例给出若干较详细的例子。  The following examples give a few more detailed examples.
实施例 3.1染料类或 /和补骨脂素类光敏剂病毒灭活剂去除滤器 Example 3.1 Dye or / and psoralen photosensitizer virus inactivating agent removal filter
本实施例 (包括 3.1.1和 3丄 2)制备的装置,其适于染料类或 /和补骨脂素类光 敏剂病毒灭活剂的除去。 本实施例中, 所用光敏剂病毒灭活剂分别为亚甲兰、 酞菁染料、 金丝桃素、 花青染料和补骨脂素。  The apparatus prepared in this example (including 3.1.1 and 3丄 2) is suitable for the removal of dyes or/and psoralen-type photosensitizer virus inactivating agents. In this embodiment, the photosensitizer virus inactivating agents used are methylene blue, phthalocyanine dye, hypericin, cyanine dye and psoralen.
实施例 3.1.1 含光敏剂病毒灭活剂吸附纤维的病毒灭活剂去除滤器 Example 3.1.1 Virus inactivating agent removal filter containing photosensitizer virus inactivating agent adsorption fiber
本实施例中, 病毒灭活剂吸附纤维为含染料类或 /和补骨脂素类光敏剂病毒 灭活剂的有机高分子物理吸附剂的纤维, 例如超细聚丙烯吸油纤维或 /和聚氨基 甲酸酯 (polyurethane) 纤维。 纤维装填厚度为 5cm。  In this embodiment, the virus inactivating agent adsorbing fiber is a fiber of an organic polymer physical adsorbent containing a dye or/and a psoralen photosensitizer virus inactivating agent, such as ultrafine polypropylene oil absorbing fiber or/and poly Polyurethane fiber. The fiber loading thickness is 5 cm.
实施例 3.1.2含光敏剂病毒灭活剂吸附纤维和吸附颗粒的病毒灭活剂去除滤器 本实施例中, 吸附颗粒为含染料类或 /和补骨脂素类光敏剂病毒灭活剂的颗 粒,例如聚丙烯基质的树脂 (例如 HP2MG系列)、含苯乙烯的大孔吸附树脂 (例如 Amberlite XAD- 7HP AmberliteXAD-16) 、 等等。 Embodiment 3.1.2 Virus Inactivating Agent Removal Filter Containing Photosensitizer Virus Inactivating Agent Adsorbing Fiber and Adsorbing Particles In this embodiment, the adsorbing particles are dye-containing or/and psoralen-based photosensitizer virus inactivating agents. Granules such as polypropylene matrix resins (e.g., HP2MG series), styrene containing macroporous adsorption resins (e.g., Amberlite XAD-7HP Amberlite XAD-16), and the like.
本实施例中,所用纤维含于所用滤材中 ,例如多种德国 Seitz公司产品 (KS80、 K900、 Supradur 80 > P30、 Eco 1000、 Permadur 0/400A, T2600、 Bio20、 Bio40、 Bio60、 Supradur 100、 等等)。 这些滤板对光敏剂病毒灭活剂、 特别是染料类光 敏剂有吸附作用。滤器直径为 1.5cm, 大孔吸附颗粒装填厚度为 3cm, 吸附颗粒 下方装填一片厚 3.5mm的滤板。 这些滤板除可进一步去除病毒灭活剂以外, 还 可防止大孔吸附颗粒脱落污染生物液体产生副作用。  In this embodiment, the fibers used are contained in the filter material used, such as various Seitz products (KS80, K900, Supradur 80 > P30, Eco 1000, Permadur 0/400A, T2600, Bio20, Bio40, Bio60, Supradur 100, and many more). These filter plates have an adsorption effect on photosensitizer virus inactivating agents, particularly dye-based photosensitizers. The filter has a diameter of 1.5 cm, the macroporous adsorption particles are packed to a thickness of 3 cm, and a 3.5 mm thick filter plate is loaded under the adsorbed particles. In addition to further removing the virus inactivating agent, these filter plates prevent the macroporous adsorption particles from falling off and contaminating the biological fluid to cause side effects.
实施例 3.2染料类病毒灭活剂去除滤器 Example 3.2 Dye-like virus inactivating agent removal filter
本实施例 (包括 3.2.1-3.2.4)制备的装置, 其适于除去染料类光敏剂病毒灭活 剂, 例如亚甲兰。  A device prepared in accordance with this embodiment (including 3.2.1-3.2.4) is suitable for removing a dye-based photosensitizer virus inactivating agent such as methylene blue.
实施例 3.2.1含聚烯烃纤维滤板的病毒灭活剂去除滤器 Example 3.2.1 Virus inactivating agent removal filter containing polyolefin fiber filter plate
本实施例中, 病毒灭活剂吸附介质选自含染料类光敏剂病毒灭活剂的有机 高分子物理吸附剂的聚烯烃纤维的滤材, 例如上述 Eco 1000、 Permadur 0/400A、 T2600、 Supradur 80> Supradur 100、Supradur 500。滤板装填 3层以上 (各厚 3.5mm)。 实施例 3.2.2含天然纤维滤板的病毒灭活剂去除滤器  In this embodiment, the virus inactivating agent adsorption medium is selected from the group consisting of a polyolefin fiber filter material of an organic polymer physical adsorbent containing a dye-based photosensitizer virus inactivating agent, such as the above-mentioned Eco 1000, Permadur 0/400A, T2600, Supradur. 80> Supradur 100, Supradur 500. The filter plate is filled with more than 3 layers (each thickness is 3.5mm). Example 3.2.2 Virus inactivating agent removal filter containing natural fiber filter plate
本实施例中, 病毒灭活剂吸附介质选自含天然纤维的滤材, 例如多种德国 Seitz公司产品 (例如, Bio20、 Bio40、 Bio60、 等等)。 滤板装填 3 层以上 (各厚 3.5mm)。  In this embodiment, the virus inactivating agent adsorption medium is selected from the group consisting of natural fiber-containing filter materials, such as various Seitz company products (e.g., Bio20, Bio40, Bio60, etc.). The filter plate is filled with more than 3 layers (each thickness is 3.5mm).
实施例 3.2.3含天然纤维的病毒灭活剂去除滤器 本实施例中, 病毒灭活剂吸附介质选自天然纤维, 例如脱脂棉纤维、 蚕丝 或木纤维。 Example 3.2.3 Virus inactivating agent removal filter containing natural fiber In this embodiment, the virus inactivating agent adsorption medium is selected from natural fibers such as cotton wool fibers, silk fibers or wood fibers.
实施例 3.2.4含合成纤维的病毒灭活剂去除滤器 Example 3.2.4 Virus inactivating agent removal filter containing synthetic fiber
本实施例中, 病毒灭活剂吸附介质选自含有机高分子物理吸附剂的合成纤 维, 例如聚氨基甲酸酯纤维或聚苯乙烯纤维。  In this embodiment, the virus inactivating agent adsorption medium is selected from the group consisting of synthetic fibers containing organic polymer physical adsorbents, such as polyurethane fibers or polystyrene fibers.
实施例 3.3染料类病毒灭活剂去除 /除白细胞滤器 Example 3.3 Dye-like virus inactivating agent removal / removal of leukocyte filter
实施例 3 中上述制备的部分装置还可用于染料类病毒灭活剂去除和除白细 胞。 实际上, 棉纤维、 聚脂、 聚氨基甲酸酯(polyurethane)、 等等, 在本实施例 中既是染料类光敏剂病毒灭活剂吸附介质、 又是去白细胞材材料。  Some of the devices prepared as described above in Example 3 can also be used for dye virus inactivating agents to remove and remove leukocytes. In fact, cotton fibers, polyesters, polyurethanes, and the like, in this embodiment, are both a dye-based photosensitizer virus inactivating agent adsorption medium and a leukocyte-removing material.
实施例 4.本发明的装置的制备 Example 4. Preparation of the device of the present invention
本实施例制备的装置, 其中的固相介质含上述有机溶剂物理吸附介质。 本实施例制备的装置, 其鉴定方法与实施例 2制备的装置的鉴定方法相同。 一般而言, 与现有的装置相比, 本实施例制备的装置有类似的病毒灭活剂除去 能力或 /和病毒灭活效率,而有明显较低的副作用 (例如与含活性炭或无机吸附介 质的装置相比): 蛋白吸附低 15%以上或 /和 APTT值升高值低 15%以上。  The apparatus prepared in this embodiment, wherein the solid phase medium contains the above organic solvent physical adsorption medium. The apparatus prepared in this example was identified in the same manner as the apparatus prepared in Example 2. In general, the devices prepared in this example have similar virus inactivating agent removal/and virus inactivation efficiencies compared to existing devices, with significantly lower side effects (eg, with activated carbon or inorganic adsorption). Comparison of medium devices): Protein adsorption is 15% lower or / and APTT value is lower than 15%.
下述实施例给出若干较详细的例子。  The following examples give a few more detailed examples.
实施例 4.1有机溶剂病毒灭活剂除去装置 Example 4.1 Organic solvent virus inactivating agent removal device
本实施例装置, 是通过将上述有机溶剂物理吸附介质装入滤桶或空心柱中 而制备的。 所用有机溶剂物理吸附介质,包括纤维 (例如, 超细聚丙烯吸油纤维)、 滤板 (例如)或 /和大孔吸附树脂 (例如 Supradur 80)、 含聚苯烯的多孔颗粒 (Amberlite XAD- 7HP. AmberliteXAD-16, 等等)。  The apparatus of this embodiment is prepared by charging the above organic solvent physical adsorption medium into a filter drum or a hollow column. Organic solvent physical adsorption medium used, including fibers (for example, ultrafine polypropylene oil absorbing fibers), filter plates (for example) or/and macroporous adsorption resins (such as Supradur 80), polyphenylene-containing porous particles (Amberlite XAD-7HP) AmberliteXAD-16, etc.).
实施例 4.2有机溶剂病毒灭活装置 Example 4.2 Organic solvent virus inactivation device
本实施例装置, 一种制备方法是通过将有机溶剂病毒灭活介质装入滤桶或 空心柱中而制备的。本实施例中,有机溶剂病毒灭活介质含上述有机溶剂物理吸 附介质和固定在吸附介质上的有机溶剂病毒灭活剂。 这一方法中所用的制备方 法, 与实施例 1.2中 "将病毒灭活剂固定至吸附介质上"的方法相同, 其中: 所 用吸附介质选自上述有机溶剂物理吸附介质 (例如实施例 4.1 中所用有机溶剂物 理吸附介质);所用病毒灭活剂为 S (例如 TnBP或 β -丙内酯)或 S D (例如 ΤηΒΡ/土 温 -80、 TnBP/ Triton X100, 或 TnBP/酒精)。  The apparatus of the present embodiment, a preparation method, is prepared by charging an organic solvent virus inactivating medium into a filter drum or a hollow column. In the present embodiment, the organic solvent virus inactivating medium contains the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium. The preparation method used in this method is the same as the method of "fixing the virus inactivating agent to the adsorption medium" in Example 1.2, wherein: the adsorption medium used is selected from the above organic solvent physical adsorption medium (for example, used in Example 4.1). The organic solvent physical adsorption medium); the virus inactivating agent used is S (for example, TnBP or β-propiolactone) or SD (for example, ΤηΒΡ/earth temperature-80, TnBP/Triton X100, or TnBP/alcohol).
另一种制备方法是: 将上述有机溶剂物理吸附介质装入滤桶或空心柱中, 预洗后再用有机溶剂分散体系以优化速度 (例如 0.1-lcm/min)流过吸附介质将有 机溶剂病毒灭活剂固定上去, 然后洗出未固定的物质。 有机溶剂分散体系的制 备方法, 与实施例 1.1 中 "有机溶剂分散体系的制备"的方法相同。 有机溶剂分 散体系的例子包括适当浓度比例的下述溶液: TnBP/土温 -80溶液、 TnBP/Triton 5Q00溶液、 TnBP/酒精溶液、 等等。 Another preparation method is: loading the above organic solvent physical adsorption medium into a filter drum or a hollow column, pre-washing, and then using an organic solvent dispersion system to flow through the adsorption medium at an optimized speed (for example, 0.1-lcm/min) to remove the organic solvent. The virus inactivating agent is fixed and then the unfixed material is washed out. The preparation method of the organic solvent dispersion system is the same as the method of "preparation of the organic solvent dispersion system" in Example 1.1. Organic solvent Examples of the dispersion system include the following solutions in appropriate concentration ratios: TnBP/earth temperature-80 solution, TnBP/Triton 5Q00 solution, TnBP/alcohol solution, and the like.
上述病毒灭活介质中含有机溶剂病毒灭活剂的含量可根椐不同需要而不 同。例如,如果所处理的生物液体的体积为 250ml, 则有的装置中 TnBP的含量 大于 7.5mg, 而有的装置中 TnBP 的含量等于或小于 7.5mg、 甚至等于或小于 2.5mg。其分别对应于有机溶剂病毒灭活剂的含量大于 3 X (lOppmX所述单人份 血液组分的体积 ml)、等于或小于 3 X (lOppmX所述单人份血液组分的体积 ml)、 及等于或小于 (lOppmX所述单人份血液组分的体积 ml)。  The content of the organic solvent-inactivated medium containing the organic solvent inactivating agent may vary depending on the needs. For example, if the volume of the biological fluid to be treated is 250 ml, the TnBP content of some devices is greater than 7.5 mg, and the TnBP content of some devices is equal to or less than 7.5 mg, or even equal to or less than 2.5 mg. It corresponds to the content of the organic solvent virus inactivating agent respectively greater than 3 X (lOppmX volume of the single-part blood component ml), equal to or less than 3 X (lOppmX volume of the single-part blood component ml), And equal to or less than (lOppmX the volume of the single-part blood component ml).
实施例 4.3有机溶剂病毒灭活 /有机溶剂病毒灭活剂除去装置 Example 4.3 Organic solvent virus inactivation / organic solvent virus inactivating agent removal device
本实施例装置, 是通过将有机溶剂病毒灭活介质和有机溶剂病毒灭活剂吸 附介质分别装入滤桶或空心柱中而制备的。有两种装填方法: A).将有机溶剂病 毒灭活介质装在靠近室的入口处, 而将有机溶剂病毒灭活剂吸附介质装在靠近 出口处; B)将有机溶剂病毒灭活剂吸附介质装在靠近室的入口处和靠近出口 处, 而将有机溶剂病毒灭活介质装在其间。 此外,也可通过有机溶剂病毒灭活装 置与有机溶剂病毒灭活剂除去装置串连而成。  The apparatus of this embodiment is prepared by separately loading an organic solvent virus inactivating medium and an organic solvent virus inactivating agent adsorption medium into a filter drum or a hollow column. There are two filling methods: A). The organic solvent virus inactivation medium is placed near the inlet of the chamber, and the organic solvent virus inactivating agent adsorption medium is placed near the outlet; B) The organic solvent virus inactivating agent is adsorbed The medium is placed near the inlet of the chamber and near the outlet, and the organic solvent virus inactivating medium is placed therebetween. Alternatively, the organic solvent virus inactivating device may be connected in series with the organic solvent virus inactivating agent removing device.
本实施例中, 所用固相介质为下述之一的组合: 1).含上述有机溶剂物理吸 附介质和固定在吸附介质上的有机溶剂病毒灭活剂的有机溶剂病毒灭活介质 (参 考实施例 4.2),和上述有机溶剂物理吸附介质 (参考实施例 4.1); 2).其它有机溶剂 病毒灭活介质 (例如实施例 1 制备的有机溶剂病毒灭活介质),和上述有机溶剂物 理吸附介质 (参考实施例 4.1); 3). 含上述有机溶剂物理吸附介质和固定在吸附介 质上的有机溶剂病毒灭活剂的有机溶剂病毒灭活介质 (参考实施例 4.2),和其它有 机溶剂病毒灭活剂吸附介质 (例如实施例 1 制备的有机溶剂病毒灭活剂吸附介 质)。  In this embodiment, the solid phase medium used is a combination of one of the following: 1) an organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (refer to the implementation) Example 4.2), and the above organic solvent physical adsorption medium (refer to Example 4.1); 2). Other organic solvent virus inactivating medium (for example, the organic solvent virus inactivating medium prepared in Example 1), and the above organic solvent physical adsorption medium (Reference Example 4.1); 3). Organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and organic solvent virus inactivating agent immobilized on the adsorption medium (refer to Example 4.2), and other organic solvent viruses The active agent adsorption medium (for example, the organic solvent virus inactivating agent adsorption medium prepared in Example 1).
实施例 4.4有机溶剂病毒灭活 /有机溶剂病毒灭活剂除去 /光敏剂病毒灭活剂除去 装置 EXAMPLES 4.4 Organic solvent virus inactivation / Organic solvent virus inactivating agent removal / Photosensitizer virus inactivating agent removal device
本实施例装置, 是通过将有机溶剂病毒灭活介质、 有机溶剂病毒灭活剂吸 附介质、 和光敏剂病毒灭活剂吸附介质 (或有机溶剂病毒灭活介质和有机溶剂病 毒灭活剂 /光敏剂病毒灭活剂吸附介质)分别装入滤桶或空心柱中而制备的。有两 种装填方法: A). 将有机溶剂病毒灭活介质装在靠近室的入口处; B)将有机溶 剂病毒灭活剂介质装在靠近室的中部。 此外,也可通过有机溶剂病毒灭活装置、 有机溶剂病毒灭活剂除去装置、 与光敏剂病毒灭活剂除去装置串连而成。  The apparatus of the present embodiment is an organic solvent virus inactivating medium, an organic solvent virus inactivating agent adsorption medium, and a photosensitizer virus inactivating agent adsorption medium (or an organic solvent virus inactivating medium and an organic solvent virus inactivating agent/photosensitive). The agent virus inactivating agent adsorption medium is prepared by separately loading into a filter drum or a hollow column. There are two methods of filling: A). The organic solvent virus inactivating medium is placed near the inlet of the chamber; B) The organic solvent virus inactivating agent medium is placed near the middle of the chamber. Alternatively, it may be formed by an organic solvent virus inactivating device, an organic solvent virus inactivating agent removing device, and a photosensitizer virus inactivating agent removing device.
本实施例中, 所用固相介质为下述之一的组合: 1).含上述有机溶剂物理吸 附介质和固定在吸附介质上的有机溶剂病毒灭活剂的有机溶剂病毒灭活介质 (参 考实施例 4.2),上述有机溶剂物理吸附介质 (参考实施例 4.1),和上述光敏剂物理 吸附纤维 (参考实施例 3); 2).其它有机溶剂病毒灭活介质 (例如实施例 1制备的 有机溶剂病毒灭活介质)、 上述有机溶剂物理吸附介质 (参考实施例 4.1)、 和上述 光敏剂物理吸附纤维 (参考实施例 3)、或其它光敏剂吸附介质 (例如实施例 1制备 的光敏剂病毒灭活剂吸附介质); 3).含上述有机溶剂物理吸附介质和固定在吸 附介质上的有机溶剂病毒灭活剂的有机溶剂病毒灭活介质 (参考实施例 4.2),和其 它有机溶剂病毒灭活剂吸附介质 (例如实施例 1制备的有机溶剂病毒灭活剂吸附 介质),和光敏剂吸附介质 (:例如上述光敏剂物理吸附纤维或实施例 1 制备的光敏 剂病毒灭活剂吸附介质); 4).含上述有机溶剂物理吸附介质和固定在吸附介质上 的有机溶剂病毒灭活剂的有机溶剂病毒灭活介质 (参考实施例 4.2),和光敏剂 /有 机溶剂病毒灭活剂吸附介质 (例如, 含苯乙烯的聚合物大孔吸附树脂、 实施例 1 制备的钝化剂 /活性碳复合物、 等等)。 In this embodiment, the solid phase medium used is a combination of one of the following: 1) an organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (see Test Example 4.2), the above organic solvent physical adsorption medium (refer to Example 4.1), and the above photosensitizer physical adsorption fiber (Reference Example 3); 2). Other organic solvent virus inactivation medium (for example, prepared in Example 1) Organic solvent virus inactivating medium), the above organic solvent physical adsorption medium (refer to Example 4.1), and the above photosensitizer physical adsorption fiber (refer to Example 3), or other photosensitizer adsorption medium (for example, the photosensitizer prepared in Example 1) Virus inactivating agent adsorption medium); 3). Organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and organic solvent virus inactivating agent immobilized on the adsorption medium (refer to Example 4.2), and other organic solvent viruses Inactivator adsorption medium (for example, organic solvent virus inactivating agent adsorption medium prepared in Example 1), and photosensitizer adsorption medium (for example, the above photosensitizer physical adsorption fiber or the photosensitizer virus inactivating agent adsorption medium prepared in Example 1) 4). An organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (Reference Example 4.2) And a photosensitizer / organic solvent virus inactivating agent adsorption medium (e.g., a polymer containing styrene macroporous adsorption resin, passivating agent prepared in Example 1 / activated carbon composite, etc.).
实施例 5.本发明的装置的制备 (4) Example 5. Preparation of the device of the present invention (4)
本实施例制备的装置, 其中的固相介质为以上实施例所制备的不同装置中 的固相介质的组合。  The apparatus prepared in this example, wherein the solid phase medium is a combination of solid phase media in different apparatuses prepared in the above examples.
多种固相介质组合时, 不同固相介质间有串连关系或并联关系。 本例中仅 列出某些固相介质串连组合的例子: 吸附介质 /钝化剂复合物与病毒灭活介质的 组合、吸附介质 /钝化剂复合物与吸附介质 /钝化剂复合物的组合、吸附介质 /钝化 剂复合物与纤维滤材的组合; 等等。 固相介质的串连组合, 可在同一个容器中 进行串连, 也可通过多个容器中进行串连。 本例中, 使用在同一个容器中进行 串连的方法。 表 3列出本实施例制备的部分装置。  When a plurality of solid phase media are combined, there are a series relationship or a parallel relationship between different solid phase media. In this example, only some examples of solid-phase media in series are listed: Combination of adsorbent/passivator complex with virus inactivating media, adsorbent/passivator complex and adsorbent/passivator complex Combination, adsorption medium/passivator composite and fiber filter; The serial combination of solid phase media can be cascaded in the same container or in multiple vessels. In this case, use the method of concatenation in the same container. Table 3 lists some of the devices prepared in this example.
表 3  table 3
装置 固相介质 1 固相介质 2 功 能  Device solid phase medium 1 solid phase medium 2 function
滤器 F1 A9 (表 1) Seitz- Bio 20 除 SD剂 /除光敏剂病毒灭活剂及其 衍生物 Filter F1 A9 (Table 1) Seitz- Bio 20 In addition to SD agent / photosensitizer virus inactivating agent and its derivatives
滤器 F2 A9 (表 1) Seitz-Supradur 除 SD剂 /除光敏剂病毒灭活剂及其 Filter F2 A9 (Table 1) Seitz-Supradur In addition to SD agent / photosensitizer virus inactivating agent
100 衍生物  100 derivatives
滤器 F3 A9 (表 1) Seitz-T2600 除 SD剂 /除光敏剂病毒灭活剂及其 衍生物  Filter F3 A9 (Table 1) Seitz-T2600 In addition to SD agent / photosensitizer virus inactivating agent and its derivatives
滤器 F4 A9 (表 1) A12 (表 1) 除 SD剂 /除光敏剂病毒灭活剂及其 衍生物  Filter F4 A9 (Table 1) A12 (Table 1) Except for SD agent / photosensitizer virus inactivating agent and its derivatives
滤器 F5 A12 (表 1) A21(表 1) 除 SD剂 /除光敏剂病毒灭活剂及其 衍生物  Filter F5 A12 (Table 1) A21 (Table 1) Except for SD agent / photosensitizer virus inactivating agent and its derivatives
滤器 F6 A15 (表 1) A28(表 1) 除 SD剂 /除光敏剂病毒灭活剂及其 衍生物 滤器 F7 A20 (表 1) A28(表 1) 除 SD剂 /除光敏剂病毒灭活剂及其 衍生物 Filter F6 A15 (Table 1) A28 (Table 1) In addition to SD agent / photosensitizer virus inactivating agent and its derivatives Filter F7 A20 (Table 1) A28 (Table 1) In addition to SD agent / photosensitizer virus inactivating agent and its derivatives
滤器 F8 A23 (表 1) A24(表 1) 除 SD剂 Filter F8 A23 (Table 1) A24 (Table 1) Except for SD agent
滤器 F9 A9 (表 1) Seitz- Bio 40, 除 SD剂 /除光敏剂病毒灭活剂及其 聚脂吸油纤维 衍生物 /除白细胞 Filter F9 A9 (Table 1) Seitz- Bio 40, in addition to SD agent / photosensitizer virus inactivating agent and its polyester oil-absorbing fiber derivative / leukocyte
滤器 G1 B6(表 2) B 12(表 2) 双重病毒灭活 Filter G1 B6 (Table 2) B 12 (Table 2) Double virus inactivation
滤器 G2 B6(表 2) B 14(表 2) 双重病毒灭活 Filter G2 B6 (Table 2) B 14 (Table 2) Double virus inactivation
滤器 G4 B6(表 2) A23(表 1) 固相 SD病毒灭活 Filter G4 B6 (Table 2) A23 (Table 1) Solid phase SD virus inactivation
滤器 G5 B 12(表 2) A21(表 1) 固相 SD病毒灭活剂病毒灭活 Filter G5 B 12 (Table 2) A21 (Table 1) Solid phase SD virus inactivating agent virus inactivation
滤器 G6 B 13(表 2) A29(表 1) 固相碘病毒灭活 Filter G6 B 13 (Table 2) A29 (Table 1) Solid phase iodine virus inactivation
滤器 G7 C2 Seitz- Bio 20,聚 固相 SD病毒灭活剂病毒灭活 Filter G7 C2 Seitz- Bio 20, solid phase SD virus inactivating agent virus inactivation
丙烯吸油纤维  Propylene oil absorbing fiber
滤器 G8 C2 Seitz- Bio 40,聚 固相 SD病毒灭活剂病毒灭活 /去白 脂吸油纤维 细胞 Filter G8 C2 Seitz- Bio 40, polymeric phase SD virus inactivating agent virus inactivation / de-white lipid absorption fiber
滤器 K1 S/D-PS 吸 氨基酸钝化的 固相 SD病毒灭活 Filter K1 S/D-PS Amino acid passivated solid phase SD virus inactivation
附树脂 PS吸附树脂  With resin PS adsorption resin
本实施例中的滤器的功能和副作用与其所含固相介质的功能和副作用一致 The function and side effects of the filter in this embodiment are consistent with the functions and side effects of the solid phase medium contained therein.
(参考上述相关实施例)。 (Refer to the related embodiment above).
实施例 6.本发明的单人份血液组分试剂盒的例子 Example 6. Example of a single blood component kit of the present invention
本发明中,术语"单人份血液组分处理装置,,是指单人份血液组分为生物液体 的生物液体处理装置。 本发明的单人份血液组分处理装置, 包括单人份血浆病 毒灭活装置、单人份血小板病毒灭活装置 (例如使用补骨脂作病毒灭活剂)、等等。 基于上述装置, 还可衍生出若干其它装置。 例如, 通过与其它处理系统 (例如血 液采集与分离处理系统)相连, 可形成集成度更高的装置。  In the present invention, the term "single-part blood component processing device" refers to a biological liquid processing device in which a single-part blood component is a biological liquid. The single-part blood component processing device of the present invention includes single-part plasma Virus inactivation device, single-part platelet virus inactivation device (for example, using psoralen as a virus inactivating agent), etc. Based on the above device, several other devices may also be derived. For example, by interacting with other processing systems (eg, The blood collection and separation processing system are connected to form a more integrated device.
本实施例 (包括 6.1-6.3)制备的试剂盒,至少含单人份血液组分处理装置,且 其为上述实施例制备的一种装置。  The kit prepared in this embodiment (including 6.1 to 6.3) contains at least a single blood component treatment device, and is a device prepared in the above embodiment.
在一些情况下, 试剂盒还含病毒灭活剂加入结构, 包括液相病毒灭活剂加 入结构或固相病毒灭活剂加入结构。 液相病毒灭活剂 (例如光敏剂病毒灭活剂溶 液)加入结构的例子为: 医用聚苯乙烯塑料管、 管中的液相病毒灭活剂、 和可去 封闭的封闭结构 (例如开关或易碎的医用聚苯乙烯塑料薄片)。  In some cases, the kit also contains a viral inactivating agent addition structure, including a liquid phase virus inactivating agent addition structure or a solid phase virus inactivating agent addition structure. Examples of liquid phase viral inactivating agents (eg, photosensitizer virus inactivating agent solutions) added to the structure are: medical polystyrene plastic tubes, liquid phase virus inactivating agents in tubes, and deblockable closed structures (eg, switches or Fragile medical polystyrene plastic sheet).
在一些情况下, 试剂盒还含病毒灭活液相反应场所, 例如用于光敏剂病毒 灭活的透明塑料袋、 等等。  In some cases, the kit also contains a viral inactivation liquid reaction site, such as a clear plastic bag for the photosensitizer virus inactivation, and the like.
在一些情况下, 试剂盒还含其它结构 (例如除白细胞滤器、 洗液、 洗液加入 装置、 等等。 其中, 洗液 (例如生理盐水)用于将装置中、 特别是柱式装置或 /和 滤器式装置中的固相介质上滞留的血液组分洗出以减小血液组分损失。  In some cases, the kit also contains other structures (eg, in addition to leukocyte filters, lotions, lotion addition devices, etc.), where a lotion (eg, saline) is used to place the device, particularly a column device or / The blood components retained on the solid phase medium in the filter device are washed out to reduce loss of blood components.
本实施例中的试剂盒的功能和副作用, 与其所含装置的功能和副作用一致 (参考上述相关实施例)。试剂盒中的其它部份若有不需要的反应活性, 可按照实 际需要加入选自上述钝化剂的物质进行钝化。钝化的方式包括: 1)对所需要钝化 的部件进行钝化、 再组装为装置; 2)对组装好的装置进行钝化; 3)对所需要钝 化的部件进行钝化、 再组装为装置、 然后对组装好的装置进行再钝化。 The function and side effects of the kit in this embodiment are consistent with the functions and side effects of the device contained therein (refer to the related embodiments above). If other parts of the kit have unwanted reactivity, follow the instructions. It is necessary to pass a substance selected from the above passivating agent for passivation. The passivation methods include: 1) passivating and reassembling the parts that need to be passivated into devices; 2) passivating the assembled device; 3) passivating and reassembling the parts that need to be passivated The device is then re-passivated for the assembled device.
实施例 6.1单人份血液组分病毒灭活试剂盒 Example 6.1 Single Human Blood Component Virus Inactivation Kit
实施例 6.1.1含病毒灭活剂吸附处理系统的单人份血液组分病毒灭活试剂盒 Example 6.1.1 Single Human Blood Component Virus Inactivating Kit Containing Virus Inactivating Agent Adsorption Treatment System
本实施例制备的试剂盒, 其含病毒灭活剂吸附处理系统, 即上述实施例制 备的含病毒灭活剂吸附介质的滤器或柱。 试剂盒还含病毒灭活剂加入装置和病 毒灭活反应场所。 其工作原理为: 单人份血液组分与病毒灭活剂加入装置加入 的病毒灭活剂接触, 在病毒灭活反应场所进行病毒灭活反应, 反应完成后生物 液体以优选的流速流过含病毒灭活剂吸附介质的滤器或柱并除去病毒灭活剂或 / 和其衍生物, 如果必要还可通过洗液加入结构加入洗液将滞留的血液组分洗出。 实施例 6.1.2含病毒灭活处理系统的单人份血液组分病毒灭活试剂盒  The kit prepared in this embodiment comprises a virus inactivating agent adsorption treatment system, i.e., a filter or column containing the virus inactivating agent adsorption medium prepared in the above embodiment. The kit also contains a viral inactivating agent addition device and a virus inactivation reaction site. The working principle is as follows: the single blood component is contacted with the virus inactivating agent added by the virus inactivating agent, and the virus inactivation reaction is carried out at the virus inactivation reaction site, and the biological fluid flows at a preferred flow rate after the reaction is completed. The virus inactivating agent adsorbs the filter or column of the medium and removes the virus inactivating agent or/and its derivative, and if necessary, washes the retained blood component by adding the washing liquid to the structure. Example 6.1.2 Single Human Blood Component Virus Inactivating Kit Containing Virus Inactivation Treatment System
本实施例制备的试剂盒, 其含病毒灭活处理系统, 即上述实施例制备的含 病毒灭活介质的滤器或柱。 其工作原理为: 单人份血液组分以优选的流速流过 含病毒灭活介质的滤器或柱进行病毒灭活 (例如, 固定化 SD剂处理、 固定化碘 处理、 或固定化离子处理), 如果必要还可通过洗液加入结构加入洗液将处理系 统中滞留的血液组分洗出。  The kit prepared in this embodiment comprises a virus inactivation treatment system, i.e., a virus inactivating medium-containing filter or column prepared in the above embodiment. The working principle is as follows: a single human blood component flows through a filter or column containing a virus inactivating medium at a preferred flow rate for virus inactivation (for example, immobilized SD agent treatment, immobilized iodine treatment, or immobilized ion treatment) If necessary, the blood components retained in the treatment system can be washed out by adding the washing liquid to the structure and adding the washing liquid.
实施例 6.1.3含病毒灭活 /病毒灭活剂吸附处理系统的单人份血液组分病毒灭活 试剂盒 Example 6.1.3 Single-part blood component virus inactivating kit containing virus inactivation/viral inactivating agent adsorption treatment system
本实施例制备的试剂盒, 其含病毒灭活处理系统, 即上述实施例制备的含 病毒灭活介质和病毒灭活剂吸附介质的滤器或柱。 其工作原理为: 单人份血液 组分以优选的流速流过滤器或柱中的病毒灭活介质进行病毒灭活 (例如, 固定化 SD剂处理、固定化碘处理、或固定化离子处理:)、并流过病毒灭活剂吸附介质保 证流出滤器或柱的血液组分基本上不含病毒灭活剂。 如果必要还可通过洗液加 入结构加入洗液将处理系统中滞留的血液组分洗出。  The kit prepared in this embodiment comprises a virus inactivation treatment system, that is, a filter or column containing the virus inactivating medium and the virus inactivating agent adsorption medium prepared in the above embodiment. The working principle is as follows: The single blood component is subjected to virus inactivation by a preferred flow rate filter or a virus inactivating medium in the column (for example, immobilized SD treatment, immobilized iodine treatment, or immobilized ion treatment: And flowing through the virus inactivating agent adsorption medium to ensure that the blood component of the effluent filter or column is substantially free of viral inactivating agents. If necessary, the blood components retained in the treatment system can be washed out by adding the washing liquid to the structure.
实施例 6.1.4含双重病毒灭活处理系统的单人份血液组分病毒灭活试剂盒 Example 6.1.4 Single Human Blood Component Virus Inactivating Kit Containing Dual Virus Inactivation Treatment System
本实施例制备的试剂盒, 其含实施例 5制备的装置, 还含病毒灭活剂加入 装置和第一次病毒灭活反应场所。 其工作原理为: 单人份血液组分与病毒灭活 剂加入装置加入的病毒灭活剂接触, 在病毒灭活反应场所进行病毒灭活反应, 反应完成后生物液体以优选的流速流过实施例 5制备的滤器或柱, 进行病毒灭 活和病毒灭活剂去除, 并使流出滤器或柱的血液组分基本上不含病毒灭活剂。 如果必要还可通过洗液加入结构加入洗液将处理系统中滞留的血液组分洗出。 实施例 6.2单人份血液组分病毒灭活 /去白细胞试剂盒 本实施例制备的试剂盒, 其含去白细胞和病毒灭活剂吸附处理系统, 即上 述实施例制备的含去白细胞和病毒灭活剂吸附介质 (或含去白细胞和病毒灭活介 质、 或含去白细胞和病毒灭活介质和病毒灭活剂吸附介质)的滤器或柱。 The kit prepared in the present embodiment, which comprises the apparatus prepared in Example 5, further comprises a virus inactivating agent addition device and a first virus inactivation reaction site. The working principle is as follows: the single blood component is contacted with the virus inactivating agent added by the virus inactivating agent, and the virus inactivation reaction is carried out at the virus inactivation reaction site, and the biological liquid is flowed through at a preferred flow rate after the reaction is completed. The filter or column prepared in Example 5 was subjected to virus inactivation and virus inactivating agent removal, and the blood components exiting the filter or column were substantially free of viral inactivating agents. If necessary, the blood components retained in the treatment system can also be washed out by adding the washing liquid to the structure to add the washing liquid. Example 6.2 Single Human Blood Component Virus Inactivation/De-Leukocyte Kit The kit prepared in this embodiment, comprising a leukocyte-removing and virus inactivating agent adsorption treatment system, that is, the de-leukocyte-containing and virus inactivating agent adsorption medium prepared by the above embodiment (or containing de-white blood cells and virus inactivating medium, or A filter or column that removes white blood cells and virus inactivating media and virus inactivating media.
实施例 7.本发明的病毒灭活方法 (1) Example 7. Virus inactivation method of the present invention (1)
本实施例 (包括 7.1-7.4)的方法, 其至少包括: A).提供生物液体; B).提供处 理系统, 所述处理系统至少含固相介质和容纳所述固相介质的室, 且至少部份 所述固相介质含钝化吸附介质; C).使所述生物液体流过所述处理系统进行病毒 灭活或病毒灭活剂吸附。 本实施例中所用处理系统选自实施例 2、 5或 6制备的 含钝化吸附介质的相应装置。  The method of the present embodiment (including 7.1-7.4), comprising at least: A) providing a biological fluid; B) providing a processing system, the processing system comprising at least a solid phase medium and a chamber containing the solid phase medium, and At least a portion of the solid phase medium comprises a passivating adsorption medium; C). flowing the biological fluid through the processing system for viral inactivation or viral inactivating agent adsorption. The treatment system used in this example is selected from the corresponding apparatus comprising the passivated adsorption medium prepared in Example 2, 5 or 6.
实施例 7.1含病毒灭活剂吸附除去处理的方法 EXAMPLES 7.1 Method for adsorption removal treatment of virus inactivating agent
实施例 7.1.1单人分血液组分病毒灭活例子 Example 7.1.1 Example of inactivation of virus by single blood component
本实施例中, 所用生物液体为单人份血小板; 所用病毒灭活剂为补骨脂素 (4,-氨甲基 -4, 5, , 8-三甲基补骨脂素); 所用装置选自实施例 6制备的试剂盒, 其中含实施例 2.1制备的装置,其含结合有钝化剂的补骨脂素类病毒灭活剂吸附 介质 (例入聚酯吸油纤维、 玻璃纤维和超细聚丙烯吸油纤维)。  In this embodiment, the biological fluid used is a single-part platelet; the virus inactivating agent used is psoralen (4,-aminomethyl-4,5,8-trimethyl psoralen); a kit selected from the preparation of Example 6, which comprises the device prepared in Example 2.1, which comprises a psoralen-type virus inactivating agent adsorption medium combined with a passivating agent (for example, polyester oil absorbing fiber, glass fiber and super Fine polypropylene oil absorbing fiber).
本实施例中病毒灭活方法为: 通过病毒灭活剂加入装置将病毒灭活剂加入 生物液体中, 在病毒灭活反应场所进行病毒灭活(适当浓度的补骨脂素、 室温、 光照 30分钟),然后使生物液体进入上述装置,通过同其中的病毒灭活剂吸附介 质接触除去病毒灭活剂。  In this embodiment, the virus inactivation method is: adding the virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing virus inactivation at the virus inactivation reaction site (appropriate concentration of psoralen, room temperature, illumination 30) Minutes), then the biological fluid is introduced into the device and the virus inactivating agent is removed by contact with the virus inactivating agent adsorption medium therein.
与使用含常用吸附介质 (例如活性碳)的相似装置的方法比较,本实施例的方 法也可有效去除病毒灭活剂 (例如补骨脂素除去 90%以上), 而具有更小的副作 用。 例如, 使用含活性炭毡的装置的对照方法中, 蛋白损失量为 10-20%, 血小 板损失 7%, 血小板形貌分数下降 16%; 而使用 2.1制备的吸附滤器, 蛋白损失 量仅为 5%, 血小板损失 3%, 血小板形貌分数基本不下降。  The method of this example is also effective in removing viral inactivating agents (e.g., more than 90% of psoralen removal) with a smaller side effect, as compared to a method using a similar device containing a conventional adsorption medium (e.g., activated carbon). For example, in a control method using a device containing activated carbon felt, the protein loss was 10-20%, the platelet loss was 7%, and the platelet morphology score was decreased by 16%. However, using the adsorption filter prepared in 2.1, the protein loss was only 5%. The platelet loss was 3%, and the platelet morphology score did not decrease.
实施例 7.1.2其它生物液体的病毒灭活 Example 7.1.2 Virus inactivation of other biological fluids
本实施例中, 所用生物液体分别为: 1.多人份合并血浆、 2.含人纤维蛋白原 的人血浆分离组分 (例如冷酒精沉淀法中的组分 I或冷沉淀)、 3.含人凝血八因子 的人血浆分离组分 (例如冷沉淀 )、4,含人凝血九因子的人血浆分离组分 (例如冷沉 淀上清液的 DEAE-Sephadex洗出组分)、 5.含人凝血酶原复合物 (PCC)的人血浆 分离组分 (例如冷沉淀上清液的 DEAE-Sephadex洗出组分)、 6.含丙种球蛋白的人 血浆分离组分 (例如冷酒精沉淀法中的组分 II沉淀:)、 7.重组 Alpha干扰素(干扰 素浓度 115万 IU/ml, pH7.0)、 8.阳性人血清参照物。  In this embodiment, the biological fluids used are: 1. multi-part combined plasma, 2. human plasma fraction containing human fibrinogen (for example, component I or cryoprecipitate in cold alcohol precipitation), 3. Human plasma separation component (eg, cryoprecipitate) containing human coagulation factor VIII, 4, human plasma separation component containing human coagulation factor IX (eg, DEAE-Sephadex washout component of cryoprecipitate supernatant), 5. Human plasma separation component of human prothrombin complex (PCC) (eg DEAE-Sephadex washout component of cryoprecipitate supernatant), 6. Human plasma fraction containing gamma globulin (eg cold alcohol precipitation method) Component II precipitation:), 7. Recombinant Alpha interferon (interferon concentration 1.15 million IU/ml, pH 7.0), 8. Positive human serum reference.
本实施例中, 所用病毒灭活剂为 S/D剂 (例如, TnBP/Triton X100、 TnBP/吐 温 80、或乙醚 /Triton X100);所用装置选自实施例 2.1制备的吸附滤器 (其尺寸根 椐生物液体的数量的不同而不同), 吸附滤器中含固定有钝化剂的有机溶剂病毒 灭活剂吸附介质 (例如含苯乙烯的聚合物大孔吸附树脂或活性碳滤板)。 In this embodiment, the virus inactivating agent used is an S/D agent (for example, TnBP/Triton X100, TnBP/spit Temperature 80, or diethyl ether / Triton X100); the apparatus used is selected from the adsorption filter prepared in Example 2.1 (the size of which varies depending on the amount of biological liquid), and the adsorption filter contains an organic solvent virus with a passivating agent immobilized thereon. Living agent adsorption medium (for example, styrene-containing polymer macroporous adsorption resin or activated carbon filter plate).
本实施例中的病毒灭活方法为: 通过病毒灭活剂加入装置将病毒灭活剂加 入生物液体中, 在病毒灭活反应场所进行病毒灭活(0.3-1% S、 或 0.3-1% S和 0.3-1% D; 30°C; 4小时), 然后使生物液体进入上述装置, 通过同其中的病毒 灭活剂吸附介质接触除去有机溶剂病毒灭活剂。  The virus inactivation method in this embodiment is: adding the virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing virus inactivation at the virus inactivation reaction site (0.3-1% S, or 0.3-1%) S and 0.3-1% D; 30 ° C; 4 hours), then the biological liquid is introduced into the above device, and the organic solvent virus inactivating agent is removed by contact with the virus inactivating agent adsorption medium therein.
与使用含常用吸附介质 (:例如活性碳)的相似装置的方法比较,本实施例的方 法也可有效去除病毒灭活剂 (例如 TnBP除去 90%以上), 而副作用则要小得多。 例如, 与使用活性碳的现有方法相比, 本实施例中的病毒灭活方法使得血浆 APTT值的增加要少 30%以上,使得其它生物液体的生物活性 (例如纤维蛋白原、 凝血 VIII因子、 凝血 IX因子、 蛋白质总量)的损失率要小 30%以上。  The method of this example is also effective in removing viral inactivating agents (e.g., more than 90% of TnBP removal) compared to methods using similar devices containing conventional adsorption media (e.g., activated carbon), with side effects being much smaller. For example, the virus inactivation method in this example results in an increase in plasma APTT value of less than 30% compared to existing methods using activated carbon, such that the biological activity of other biological fluids (eg, fibrinogen, coagulation factor VIII) The loss rate of coagulation factor IX and total protein is 30% or less.
实施例 7.2含固定化病毒灭活剂病毒灭活处理的方法 EXAMPLES 7.2 Method for inactivating treatment with immobilized virus inactivating agent virus
实施例 7.2.1单人分血液组分的处理 Example 7.2.1 Treatment of single blood component
本实施例中, 所用生物液体为单人份血浆; 所用装置选自实施例 6制备的 试剂盒,其中含实施例 2.2制备的病毒灭活装置,其分别含有机溶剂病毒灭活剂 /吸附介质 /钝化剂复合物、 Crnio-Zetaplus VR灭病毒滤板和 Seitz-碘 -PVPP滤板。 需要强调的是,本实施例中的病毒灭活处理,可以使血浆制品厂在采浆后即进行 一次病毒灭活,以减小在血 贮、运和生产过程中的病毒污染风险,提高产品病毒 安全性。  In the present embodiment, the biological fluid used is a single-part plasma; the apparatus used is selected from the kit prepared in Example 6, which comprises the virus inactivating apparatus prepared in Example 2.2, which respectively contains the organic solvent virus inactivating agent/adsorption medium. / Passivator complex, Crnio-Zetaplus VR virus filter plate and Seitz-iodine-PVPP filter plate. It should be emphasized that the virus inactivation treatment in the present embodiment can enable the plasma product factory to perform virus inactivation after the pulping to reduce the risk of virus contamination in the blood storage, transportation and production process, and improve the product. Virus security.
本实施例中病毒灭活方法为: 将室温下的生物液体以优选的流速 (例如线速 度 0.1-1.5cm/分:)流过上述装置,进行病毒灭活。如有必要,用冼液洗涤包括固相 介质在内的装置。  The virus inactivation method in this embodiment is: a biological fluid at room temperature is passed through the above apparatus at a preferred flow rate (e.g., a linear velocity of 0.1 - 1.5 cm / min:) to carry out virus inactivation. If necessary, wash the device including the solid phase media with sputum.
与使用含常用吸附介质 (:例如活性碳)的相似装置的方法比较,本实施例的方 法可同等有效灭活病毒 (例如模式病毒灭活 4个 log以上), 而具有更小的不需要 的反应活性 (例如, APTT值的增加要少 20%以上, 纤维蛋白原、凝血 VIII因子、 凝血 IX因子、 蛋白质总量的损失率要小 15%以上)。  Compared with the method using a similar device containing a common adsorption medium (for example, activated carbon), the method of the present embodiment can equally effectively inactivate the virus (for example, the pattern virus is inactivated by more than 4 logs), and has less unnecessary Reactivity (for example, the APTT value is increased by more than 20%, and the loss rate of fibrinogen, coagulation factor VIII, coagulation factor IX, and total protein is 15% or less).
实施例 7.2.2其它生物液体的处理  Example 7.2.2 Treatment of other biological fluids
本实施例中, 所用生物液体分别为: 1.多人份合并血浆、 2.含人纤维蛋白原 的人血浆分离组分 (例如冷酒精沉淀法中的组分 I或冷沉淀)、 3含人凝血酶原复 合物 (PCC)的人血浆分离组分 (例如冷沉淀上清液的 DEAE-Sephadex洗出组分)。 本实施例中, 所用装置选自实施例 2.2制备的病毒灭活装置 (室的体积根椐生物 液体体积不同而不同), 其分别含吸附介质 /有机溶剂病毒灭活剂 /钝化剂复合物、 Cuno-Zetaplus VR灭病毒滤板和 Seitz-碘 -PVPP滤板。 In this embodiment, the biological fluids used are: 1. multi-part combined plasma, 2. human plasma fraction containing human fibrinogen (for example, component I or cryoprecipitate in cold alcohol precipitation), 3 Human plasma separation component of human prothrombin complex (PCC) (eg, DEAE-Sephadex washout component of cryoprecipitate supernatant). In this embodiment, the apparatus used is selected from the virus inactivation apparatus prepared in Example 2.2 (the volume of the chamber is different from the volume of the biological liquid), and the adsorbent medium/organic solvent virus inactivating agent/passivator complex is respectively contained. , Cuno-Zetaplus VR virus-killing filter plate and Seitz-iodine-PVPP filter plate.
本实施例中病毒灭活方法与实施例 7.2.1中的方法相同, 其与使用含常用吸 附介质 (例如活性碳)的相似装置的方法的比较亦与实施例 7.2.1中的比较一致。 实施例 7.3含去除病毒灭活剂和白细胞步骤的方法  The virus inactivation method in this example is the same as in the method of Example 7.2.1, and the comparison with the method using a similar apparatus containing a usual adsorption medium (e.g., activated carbon) is also in agreement with the comparison in Example 7.2.1. EXAMPLE 7.3 Method for removing steps of virus inactivating agent and leukocyte
本实施例中, 所用生物液体为单人份血浆; 所用病毒灭活剂选自光敏剂病 毒灭活剂 (例如, 4,-氨甲基 -4, 5,, 8-三甲基补骨脂素; 所用试剂盒选自实施例 7制备的试剂盒, 其中含实施例 2.4制备的去白细胞 /病毒灭活剂去除滤器。  In this embodiment, the biological fluid used is a single-part plasma; the virus inactivating agent used is selected from the photosensitizer virus inactivating agent (for example, 4,-aminomethyl-4,5,8-trimethyl psoralen) The kit used was selected from the kit prepared in Example 7, which contained the leukocyte/virus inactivating agent removal filter prepared in Example 2.4.
本实施例中的白细胞去除 /病毒灭活剂去除方法为: 通过病毒灭活剂加入装 置将病毒灭活剂加入生物液体中,在病毒灭活反应场所进行病毒灭活(例如,适 当浓度的病毒灭活剂、室温、光照 30分钟), 然后使生物液体进入上述装置, 通 过同其中的固相介质接触除去病毒灭活剂和其衍生物。  The leukocyte removal/viral inactivating agent removal method in this embodiment is: adding a virus inactivating agent to a biological liquid through a virus inactivating agent addition device, and performing virus inactivation at a virus inactivation reaction site (for example, an appropriate concentration of virus) The inactivating agent, room temperature, light for 30 minutes), then the biological liquid is introduced into the above device, and the virus inactivating agent and its derivative are removed by contact with the solid phase medium therein.
与使用含常用吸附介质 (例如玻璃纤维)的相似装置的方法比较,本实施例的 方法有同等的白细胞去除效率, 而副作用则明显降低。 实施例 8.本发明的病毒灭活方法 (2)  The method of this example has an equivalent leukocyte removal efficiency and a significant reduction in side effects as compared to a method using a similar device containing a conventional adsorption medium such as glass fiber. Example 8. Virus inactivation method of the present invention (2)
本实施例 (包括 8.1-8.3)的方法, 其至少包括: A).提供生物液体; B).使所述 生物液体与病毒灭活剂接触并进行病毒灭活 (例如, 适当浓度的病毒灭活剂、 室 温、 光照 30分钟), 其中所述病毒灭活剂包括光敏剂病毒灭活剂; C).提供处理 系统, 所述处理系统至少含病毒灭活剂吸附介质和容纳所述固相介质的室, 其 中所述病毒灭活剂吸附介质含光敏剂物理吸附纤维; D).使经 B)步骤处理的生物 液体流过所述处理系统去除光敏剂病毒灭活剂吸附及其衍生物。  The method of the present embodiment (including 8.1-8.3), comprising at least: A) providing a biological fluid; B) contacting the biological fluid with a viral inactivating agent and performing virus inactivation (eg, an appropriate concentration of virus is eliminated) Living agent, room temperature, illumination for 30 minutes), wherein the virus inactivating agent comprises a photosensitizer virus inactivating agent; C) providing a treatment system, the treatment system containing at least a virus inactivating agent adsorption medium and containing the solid phase a chamber of the medium, wherein the virus inactivating agent adsorption medium contains a photosensitizer physical adsorption fiber; D). flowing the biological liquid treated by the step B) through the treatment system to remove the photosensitizer virus inactivating agent adsorption and its derivative .
本实施例中, 所用生物液体为单人份血浆; 所用处理系统为实施例 7制备 的装置,其含选自实施例 3制备的装置 (滤器或柱)。  In the present embodiment, the biological fluid used was a single-part plasma; the treatment system used was the apparatus prepared in Example 7, which contained a device (filter or column) selected from the preparation of Example 3.
与使用含常用吸附介质 (例如活性碳)的相似装置的方法比较,本实施例的方 法可有效去除灭活病毒剂 (例如 95%以上), 而具有更小的不需要的反应活性 (例 如, APTT值的增加要少 20%以上, 纤维蛋白原、凝血 VIII因子、凝血 IX因子、 蛋白质总量的损失率要小 15%以上)。  The method of the present embodiment can effectively remove inactivated virus agents (e.g., more than 95%) with less unwanted reactivity (e.g., compared to methods using similar devices containing conventional adsorption media (e.g., activated carbon). The APTT value is increased by more than 20%, and the loss rate of fibrinogen, coagulation factor VIII, coagulation factor IX, and total protein is 15% or less).
实施例 8.1含染料类或 /和补骨脂素类光敏剂病毒灭活剂去除步骤的方法 EXAMPLES 8.1 Method for removing a dye-containing or/and psoralen-based photosensitizer virus inactivating agent
本实施例中, 所用病毒灭活剂选自染料类或补骨脂素类光敏剂病毒灭活剂 (例如, 亚甲兰或 4,-氨甲基 -4, 5,, 8-三甲基补骨脂素:); 所用装置选自实施例 6 制备的试剂盒, 其中含实施例 3.1制备的吸附滤器。  In this embodiment, the virus inactivating agent used is selected from the group consisting of dyes or psoralen photosensitizer virus inactivating agents (for example, methylene blue or 4,-aminomethyl-4,5,8-trimethyl) Psoralen:) The device used was selected from the kit prepared in Example 6, which contained the adsorption filter prepared in Example 3.1.
实施例 8.2含染料类光敏剂病毒灭活剂去除步骤的方法  EXAMPLES 8.2 Method for removing step of dye-containing photosensitizer virus inactivating agent
本实施例中,所用病毒灭活剂选自染料类光敏剂病毒灭活剂(例如亚甲兰); 所用装置选自实施例 6制备的试剂盒, 其中含实施例 3.2制备的吸附滤器。 In this embodiment, the virus inactivating agent used is selected from the group consisting of a dye-based photosensitizer virus inactivating agent (for example, methylene blue); The apparatus used was selected from the kit prepared in Example 6, which contained the adsorption filter prepared in Example 3.2.
实施例 8.3含染料类光敏剂病毒灭活剂去除 /除白细胞步骤的方法 EXAMPLES 8.3 Method for removing/removing leukocyte-containing step of dye-containing photosensitizer virus inactivating agent
本实施例中,所用病毒灭活剂选自染料类光敏剂病毒灭活剂(例如亚甲兰); 所用装置选自实施例 6制备的试剂盒, 其中含实施例 3.3制备的吸附滤器。  In the present embodiment, the virus inactivating agent used is selected from a dye-based photosensitizer virus inactivating agent (e.g., methylene blue); and the apparatus used is selected from the kit prepared in Example 6, which contains the adsorption filter prepared in Example 3.3.
实施例 9.本发明的病毒灭活方法 (3) Example 9. Virus inactivation method of the present invention (3)
本实施例 (包括 9.1-9.3)中的病毒灭活方法至少包括: A).提供生物液体; B). 提供处理系统, 所述处理系统至少含固相介质和容纳所述固相介质的室, 其中 所述固相介质包括病毒灭活介质或 /和病毒灭活剂吸附介质, 且:所述病毒灭活介 质至少含有机溶剂物理吸附介质和固定在其上的有机溶剂病毒灭活剂, 所述病 毒灭活剂吸附介质至少包括有机溶剂物理吸附介质; C).使所述生物液体流过所 述处理系统进行病毒灭活或 /和病毒灭活剂吸附。 本实施例中所用处理系统选自 实施例 4制备的装置。  The virus inactivation method in this embodiment (including 9.1-9.3) includes at least: A) providing a biological fluid; B) providing a treatment system, the treatment system comprising at least a solid phase medium and a chamber containing the solid phase medium Wherein the solid phase medium comprises a virus inactivating medium or/and a virus inactivating agent adsorption medium, and: the virus inactivating medium comprises at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon, The virus inactivating agent adsorption medium comprises at least an organic solvent physical adsorption medium; C). flowing the biological liquid through the treatment system for virus inactivation or/and virus inactivating agent adsorption. The treatment system used in this example was selected from the apparatus prepared in Example 4.
实施例 9.1含有机溶剂病毒灭活剂吸附除去步骤的方法 EXAMPLES 9.1 Method for adsorption removal step of organic solvent-containing virus inactivating agent
实施例 9.1.1单人分血液组分病毒灭活例子 Example 9.1.1 Example of inactivation of virus by single blood component
本实施例中, 所用生物液体为单人份血浆; 所用装置选自实施例 6制备的 试剂盒, 其中含实施例 4.1制备的装置。  In the present embodiment, the biological fluid used was a single-part plasma; the apparatus used was selected from the kit prepared in Example 6, which contained the apparatus prepared in Example 4.1.
本实施例中的病毒灭活方法与实施例 7.1.1中病毒灭活方法相同, 其病毒灭 活剂除去效率和副作用降低亦相同。  The virus inactivation method in this example is the same as the virus inactivation method in Example 7.1.1, and the virus inactivating agent removal efficiency and side effect reduction are also the same.
实施例 9.1.2其它生物液体的病毒灭活例子 Example 9.1.2 Examples of virus inactivation of other biological fluids
本实施例中, 所用生物液体与实施例 7.2.2中所用生物液体相同; 所用装置 选自实施例 4.1制备的滤器或柱 (室的体积根椐生物液体体积不同而不同)。 本实 施例中的病毒灭活方法与实施例 7.1.2中病毒灭活方法相同, 其病毒灭活剂除去 效率和副作用降低亦相同。  In the present embodiment, the biological fluid used is the same as that used in the embodiment 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.1 (the volume of the chamber varies depending on the volume of the biological fluid). The virus inactivation method in this embodiment is the same as the virus inactivation method in Example 7.1.2, and the virus inactivating agent removal efficiency and side effect reduction are also the same.
实施例 9.2含固定化有机溶剂病毒灭活步骤的方法 EXAMPLES 9.2 Method for Inactivating Step of Immobilized Organic Solvent Virus
实施例 9.2.1单人分血液组分病毒灭活例子 Example 9.2.1 Example of inactivation of virus by single blood component
本实施例中, 所用生物液体为单人份血浆; 所用装置选自实施例 6制备的 试剂盒, 其中含实施例 4.2制备的装置。 本实施例中的病毒灭活方法与实施例 7.2.1中病毒灭活方法相同, 其病毒灭活效率和副作用降低亦相同。  In the present embodiment, the biological fluid used was a single-part plasma; the apparatus used was selected from the kit prepared in Example 6, which contained the apparatus prepared in Example 4.2. The virus inactivation method in this example is the same as the virus inactivation method in Example 7.2.1, and the virus inactivation efficiency and side effect reduction are also the same.
实施例 9.2.2其它生物液体的病毒灭活例子 Example 9.2.2 Examples of virus inactivation of other biological fluids
本实施例中, 所用生物液体与实施例 7.2.2中所用生物液体相同; 所用装置 选自实施例 4.2制备的滤器或柱 (室的体积根椐生物液体体积不同而不同)。 本实 施例中的病毒灭活方法与实施例 7.2.2中病毒灭活方法相同, 其病毒灭活效率和 副作用降低亦相同。 实施例 9.3含固定化有机溶剂病毒灭活 /有机溶剂病毒灭活剂除去步骤的方法 实施例 9.3.1单人分血液组分病毒灭活例子 In the present embodiment, the biological fluid used is the same as the biological fluid used in Example 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.2 (the volume of the chamber is different from the volume of the biological fluid). The virus inactivation method in this embodiment is the same as the virus inactivation method in Example 7.2.2, and the virus inactivation efficiency and side effect reduction are also the same. Example 9.3 Method for Removal of Immobilized Organic Solvent Virus Inactivation/Organic Solvent Virus Inactivating Agent Example 9.3.1 Single Human Blood Component Virus Inactivation Example
本实施例中, 所用生物液体为单人份血浆; 所用装置选自实施例 6制备的 试剂盒, 其中含实施例 4.3制备的滤器或柱。  In the present embodiment, the biological fluid used is a single-part plasma; the apparatus used is selected from the kit prepared in Example 6, which contains the filter or column prepared in Example 4.3.
本实施例中病毒灭活方法为: 将室温下的生物液体以优选的流速流过上述 装置, 进行病毒灭活并除去可能脱落的有机溶剂病毒灭活剂。  The virus inactivation method in this embodiment is: flowing the biological fluid at room temperature through the above device at a preferred flow rate, performing virus inactivation and removing the organic solvent virus inactivating agent which may fall off.
本实施例方法中的病毒灭活效率和副作用与实施例 9.2.2方法相同, 从所用 装置出口收集的生物液体中的有机溶剂含量小于 10ppm。  The virus inactivation efficiency and side effects in the method of this example were the same as in the method of Example 9.2.2, and the content of the organic solvent in the biological liquid collected from the outlet of the apparatus used was less than 10 ppm.
实施例 9.3.2其它生物液体的病毒灭活例子 Example 9.3.2 Examples of virus inactivation of other biological fluids
本实施例中, 所用生物液体与实施例 7.2.2中所用生物液体相同; 所用装置 选自实施例 4.3制备的滤器或柱 (室的体积根椐生物液体体积不同而不同)。  In the present embodiment, the biological fluid used is the same as that used in the embodiment 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.3 (the volume of the chamber varies depending on the volume of the biological fluid).
本实施例中病毒灭活方法为: 将室温下的生物液体以优选的流速流过上述 装置, 进行病毒灭活并除去可能脱落的有机溶剂病毒灭活剂。 本实施例中方法 的病毒灭活效率、副作用和收集的生物液体中的有机溶剂含量与实施例 9.3.1方 法相一致。  The virus inactivation method in this embodiment is: flowing the biological fluid at room temperature through the above device at a preferred flow rate, performing virus inactivation and removing the organic solvent virus inactivating agent which may fall off. The virus inactivation efficiency, side effects, and organic solvent content of the collected biological fluids in the method of this example were consistent with the method of Example 9.3.1.
实施例 10.本发明的病毒灭活方法 (4) Example 10. Viral inactivation method of the present invention (4)
本实施例 (包括 10.1和 10.2)中的病毒灭活方法至少包括: A).提供生物液体; B).对所述生物液体进行第一次病毒灭活; C).提供处理系统, 所述处理系统至少 含病毒灭活介质和容纳所述固相介质的室, 且其中所述病毒灭活介质至少含有 机溶剂物理吸附介质和固定在其上的有机溶剂病毒灭活剂; D).使经第一次病毒 灭活的所述生物液体流过所述处理系统进行第二次病毒灭活。 本实施例中, 所 用生物液体为单人份血桨。 本实施例中, 所用装置选自实施例 6制备的试剂盒, 其中含选自实施例 4.4或实施例 5制备的装置。  The method of virus inactivation in this embodiment (including 10.1 and 10.2) comprises at least: A) providing a biological fluid; B) performing a first viral inactivation of the biological fluid; C) providing a treatment system, The treatment system comprises at least a virus inactivating medium and a chamber containing the solid phase medium, and wherein the virus inactivating medium contains at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon; D). The biological fluid inactivated by the first virus flows through the treatment system for a second viral inactivation. In this embodiment, the biological fluid used is a single blood paddle. In this embodiment, the apparatus used is selected from the kit prepared in Example 6, which comprises a device selected from the preparation of Example 4.4 or Example 5.
本实施例中, 使用两种不同的病毒灭活机理, 病毒灭活效率较高。  In this embodiment, two different virus inactivation mechanisms are used, and the virus inactivation efficiency is high.
实施例 10.1含光敏剂病毒灭活和有机溶剂病毒灭活步骤的病毒灭活 EXAMPLES 10.1 Virus Inactivation of Photosensitizer Virus Inactivation and Organic Solvent Virus Inactivation Steps
本实施例中, 所用光敏剂病毒灭活剂选自染料类或补骨脂素类光敏剂病毒 灭活剂(例如, 亚甲兰或 4'-氨甲基 -4, 5,, 8-三甲基补骨脂素)。 本实施例中, 所用处理糸统选自实施例 4.4或实施例 5制备的装置,特别是含光敏剂病毒灭活 剂吸附介质、 有机溶剂病毒灭活介质和有机溶剂病毒灭活剂吸附介质,其中所述 病毒灭活介质至少含有机溶剂物理吸附介质和固定在其上的有机溶剂病毒灭活 剂。  In this embodiment, the photosensitizer virus inactivating agent used is selected from the group consisting of dyes or psoralen photosensitizer virus inactivating agents (for example, methylene blue or 4'-aminomethyl-4, 5, 8-3 Methyl psoralen). In this embodiment, the treatment system used is selected from the apparatus prepared in Example 4.4 or Example 5, in particular, a photosensitizer virus inactivating agent adsorption medium, an organic solvent virus inactivating medium, and an organic solvent virus inactivating agent adsorption medium. Wherein the virus inactivating medium comprises at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon.
本实施例中,病毒灭活方法为: 通过病毒灭活剂加入装置将光敏剂病毒灭活 剂加入生物液体中,在病毒灭活反应场所进行第一次病毒灭活(例如,适当浓度 的病毒灭活剂、室温、 光照 30分钟), 然后使生物液体进入上述装置, 通过同其 中的固相介质接触除去光敏剂病毒灭活剂并进行第二次病毒灭活、 并除去光敏 剂病毒灭活剂和有机溶剂病毒灭活剂。 In this embodiment, the virus inactivation method is: adding the photosensitizer virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing the first virus inactivation at the virus inactivation reaction site (for example, an appropriate concentration) Virus inactivating agent, room temperature, light for 30 minutes), then let the biological fluid enter the above device, remove the photosensitizer virus inactivating agent by contact with the solid phase medium therein and carry out the second virus inactivation, and remove the photosensitizer virus Inactivating agent and organic solvent virus inactivating agent.
本实施例中, 由于使用副作用较低的固相介质, 副作用控制在可以接受的 水平。 例如, 单人分血浆经上述病毒灭活处理后, APTT值保持在 47秒以内, 纤维蛋白原、 凝血 VIII因子、 凝血 IX因子的损失率也分别为小于 30%。  In this embodiment, the side effects are controlled at an acceptable level due to the use of a solid phase medium having a lower side effect. For example, after the single-person plasma is inactivated by the above virus, the APTT value is kept within 47 seconds, and the loss rates of fibrinogen, coagulation factor VIII, and coagulation factor IX are also less than 30%, respectively.
实施例 10.2含病毒热灭活和有机溶剂病毒灭活步骤的病毒灭活 Example 10.2 Virus inactivation with virus heat inactivation and organic solvent virus inactivation steps
本实施例中, 所用装置选自实施例 6制备的试剂盒, 其含实施例 2.2或 4.3 制备的滤器或柱, 其含有机溶剂病毒灭活介质和有机溶剂病毒灭活剂吸附介质。  In the present embodiment, the apparatus used is selected from the kit prepared in Example 6, which comprises the filter or column prepared in Example 2.2 or 4.3, which contains an organic solvent virus inactivating medium and an organic solvent virus inactivating agent adsorption medium.
本实施例中病毒灭活方法为: 通过保护剂加入装置将公知的血浆活性保护剂 (例如糖类、 枸缘酸钠、 钙、 等等)加入生物液体中, 在病毒灭活反应场所进行病 毒灭活(例如, 适当浓度的保护剂、 52° C、 3 小时), 然后使生物液体进入上述 装置, 通过同其中的固相介质接触进行有机溶剂病毒灭活及除去可能脱落的有 机溶剂病毒灭活剂。  The method for inactivating the virus in this embodiment is: adding a known plasma active protective agent (for example, sugar, sodium valerate, calcium, etc.) to the biological liquid through the protective agent adding device, and performing virus at the virus inactivation reaction site. Inactivate (for example, a suitable concentration of protective agent, 52 ° C, 3 hours), then let the biological liquid enter the above device, inactivate the organic solvent virus by contact with the solid phase medium and remove the organic solvent that may fall off Active agent.
本实施例中, 由于使用副作用较低的固相介质, 至少第二次病毒灭活的副 作用是较小的。  In this embodiment, the side effect of at least the second virus inactivation is small due to the use of a solid phase medium having a lower side effect.
实施例 11.本发明的病毒灭活方法 (5) Example 11. Virus inactivation method of the present invention (5)
本实施例中的病毒灭活方法至少包括: A).提供生物液体; B).提供实施例 2 制备的装置, 其含所述病毒灭活介质或 /和病毒灭活剂吸附介质; C).使所述生物 液体流过所述处理系统进行病毒灭活或 /和病毒灭活剂吸附、 以及白细胞去除。  The virus inactivation method in this embodiment at least comprises: A) providing a biological fluid; B) providing the device prepared in Example 2, comprising the virus inactivating medium or/and a virus inactivating agent adsorption medium; The biological fluid is passed through the treatment system for virus inactivation or/and viral inactivating agent adsorption, and leukocyte removal.
应当清楚, 对于本领域的技术人员来说, 对这里所述的本发明的优选的实 施方案显然可以作出各种变动和修改。 在不偏离本发明的精神和范围及不减小 其优点的情况下, 可以进行这些修改和变动。 因此, 这些修改和变动包括在所 附的权利要求的范围内。  It will be apparent that various modifications and changes can be made to the preferred embodiments of the invention described herein. These modifications and variations can be made without departing from the spirit and scope of the invention and the advantages thereof. Accordingly, such modifications and variations are intended to be included within the scope of the appended claims.

Claims

权 利 要 求 书 Claim
1. 一种病毒灭活方法, 至少包括: A method for inactivating a virus, comprising at least:
A).提供生物液体; B).提供处理系统; 和 C).使所述生物液体与所述处理系统 接触,  A) providing a biological fluid; B) providing a treatment system; and C) contacting the biological fluid with the treatment system,
其中所述处理系统, 至少包含固相介质和容纳所述固相介质的室,  Wherein the processing system comprises at least a solid phase medium and a chamber containing the solid phase medium,
其中所述固相介质, 至少包含:  Wherein the solid phase medium comprises at least:
a) .病毒灭活剂吸附介质; 和 /或  a) a virus inactivating agent adsorption medium; and / or
b) .病毒灭活介质,所述病毒灭活介质包含病毒灭活剂吸附介质和固定在 其上的病毒灭活剂,  b) a virus inactivating medium comprising a virus inactivating agent adsorption medium and a virus inactivating agent immobilized thereon,
其中所述病毒灭活剂吸附介质, 包括:  Wherein the virus inactivating agent adsorption medium comprises:
(a) .钝化吸附介质,所述钝化吸附介质至少含吸附介质和固定在吸附介质 上的钝化剂; 或 /和  (a) passivating an adsorption medium containing at least an adsorption medium and a passivating agent immobilized on the adsorption medium; or / and
(b) .物理吸附介质, 所述物理吸附介质包括:  (b) physical adsorption medium, the physical adsorption medium comprising:
(i) .用作光敏剂病毒灭活剂吸附介质的光敏剂物理吸附纤维, 所述光 敏剂物理吸附纤维含光敏剂病毒灭活剂的有机高分子物理吸附剂; 和 (i) a photosensitizer physically adsorbing fiber used as a photosensitizer virus inactivating agent adsorption medium, said photosensitive agent physically adsorbing an organic polymer physical adsorbent containing a photosensitizer virus inactivating agent;
(ii) .用作有机溶剂病毒灭活剂吸附介质的有机溶剂物理吸附介质,所述 有机溶剂物理吸附介质含有机溶剂病毒灭活剂的有机高分子物理吸 附剂。 (ii) An organic solvent physical adsorption medium used as an organic solvent virus inactivating agent adsorption medium, the organic solvent physical adsorption medium containing an organic solvent physical adsorbent of an organic solvent virus inactivating agent.
2. 权利要求 1所述的方法, 其中所述固相介质包括有机溶剂病毒灭活剂吸附介 质,且所述有机溶剂病毒灭活剂吸附介质至少含所述钝化吸附介质,或 /和有 机溶剂物理吸附介质。  2. The method of claim 1, wherein the solid phase medium comprises an organic solvent virus inactivating agent adsorption medium, and the organic solvent virus inactivating agent adsorption medium contains at least the passivation adsorption medium, or/and organic Solvent physical adsorption medium.
3. 权利要求 1所述的方法, 其中所述固相介质包括有机溶剂病毒灭活介质, 且 所述有机溶剂病毒灭活介质至少含有机溶剂病毒灭活剂、和所述钝化吸附介 质、 或 /和有机溶剂物理吸附介质。  3. The method of claim 1, wherein the solid phase medium comprises an organic solvent virus inactivating medium, and the organic solvent virus inactivating medium comprises at least an organic solvent virus inactivating agent, and the passivating adsorption medium, Or / and organic solvent physical adsorption medium.
4. 权利要求 1-3之一所述的方法, 其中所述有机溶剂物理吸附介质包括大孔吸 附树脂或 /和纤维。  The method of any one of claims 1 to 3, wherein the organic solvent physic adsorption medium comprises macroporous resin or/and fiber.
5. 权利要求 1-4之一所述的方法, 其中所述有机溶剂病毒灭活剂的有机高分子 物理吸附剂包括聚烯烃。  The method according to any one of claims 1 to 4, wherein the organic polymer physical adsorbent of the organic solvent virus inactivating agent comprises a polyolefin.
6. 权利要求 5所述的方法, 其中所述聚烯烃包括聚苯乙烯。  6. The method of claim 5 wherein the polyolefin comprises polystyrene.
7. 权利要求 1所述的方法,其中所述固相介质包括光敏剂病毒灭活剂吸附介质, 且所述光敏剂病毒灭活剂吸附介质至少含所述钝化吸附介质,或 /和光敏剂物 理吸附纤维。  7. The method of claim 1 wherein said solid phase medium comprises a photosensitizer virus inactivating agent adsorption medium, and said photosensitizer virus inactivating agent adsorption medium comprises at least said passivated adsorption medium, or/and photosensitive The agent physically adsorbs the fiber.
8. 权利要求 1或 7之一所述的方法, 其中所述光敏剂物理吸附纤维包括天然纤 维。 8. The method of any of claims 1 or 7, wherein the photosensitizer physico-adsorbing fibers comprise natural fibers.
9. 权利要求 8所述的方法, 其中所述天然纤维包括木纤维。 9. The method of claim 8 wherein the natural fibers comprise wood fibers.
10.权利要求 1 、 7或 8所述的方法, 特征在于: 其中所述光敏剂物理吸附纤维 包括合成纤维。  10. The method of claim 1, 7 or 8, wherein: said photosensitizer physically adsorbing fibers comprise synthetic fibers.
11.权利要求 10所述的方法, 其中所述合成纤维包括聚烯烃纤维。  11. The method of claim 10, wherein the synthetic fibers comprise polyolefin fibers.
12.权利要求 1-10之一所述的方法,其中所述固相介质包含所述有机溶剂病毒灭 活介质、 有机溶剂病毒灭活剂吸附介质、 或 /和光敏剂物理吸附纤维。  The method of any one of claims 1 to 10, wherein the solid phase medium comprises the organic solvent virus inactivation medium, the organic solvent virus inactivating agent adsorption medium, or/and the photosensitizer physical adsorption fiber.
13.权利要求 1所述的方法, 其中所述固相介质包括碘病毒灭活介质, 所述碘病 毒灭活介质至少含碘、 碘吸附介质和钝化剂。  13. The method of claim 1 wherein said solid phase medium comprises an iodine virus inactivating medium, said iodine virus inactivating medium comprising at least iodine, an iodine adsorption medium and a passivating agent.
14.权利要求 1-13之一所述的方法, 其中所述固相介质包括深层过滤滤材,所述 深层过滤滤材含所述病毒灭活剂吸附介质,且具有下述特征: A).平均密度大 于 0.25g/cm3; 和 B).灰分小于 1%。 The method according to any one of claims 1 to 13, wherein the solid phase medium comprises a depth filter medium, the deep filter medium contains the virus inactivating agent adsorption medium, and has the following characteristics: A) The average density is greater than 0.25 g/cm 3 ; and B). The ash content is less than 1%.
15.权利要求 1-14之一所述的方法,其中所述钝化吸附介质中的吸附介质包括活 性炭。  The method of any one of claims 1-14, wherein the adsorbent medium in the passivating adsorption medium comprises activated carbon.
16.权利要求 1-14之一所述的方法,其中所述钝化吸附介质中的吸附介质包括离 子交换吸附介质。  16. The method of any of claims 1-14, wherein the adsorbent medium in the passivating adsorption medium comprises an ion exchange adsorption medium.
17.权利要求 1-14之一所述的方法,其中所述钝化吸附介质中的吸附介质包括所 述物理吸附介质。  The method of any one of claims 1-14, wherein the adsorbent medium in the passivating adsorption medium comprises the physical adsorption medium.
18.权利要求 1-17之一所述的方法,其中所述钝化吸附介质中的钝化剂,包括具 有亲水基团或 /和亲油基团、且可降低所述固相介质对所述生物液体的副作用 的有机物。  The method of any one of claims 1 to 17, wherein the passivating agent in the passivating adsorption medium comprises a hydrophilic group or/and an oleophilic group, and the solid phase medium pair can be lowered An organic substance that describes the side effects of biological fluids.
19.权利要求 18所述的方法, 其中所述纯化剂包括天然油脂。  19. The method of claim 18, wherein the purifying agent comprises a natural fat.
20.权利要求 18所述的方法, 其中所述纯化剂包括羟基化合物。  20. The method of claim 18, wherein the purification agent comprises a hydroxy compound.
21.权利要求 18所述的方法, 其中所述纯化剂包括氨基酸。  21. The method of claim 18, wherein the purification agent comprises an amino acid.
22.权利要求 18所述的方法, 其中所述纯化剂包括有机溶剂。  22. The method of claim 18, wherein the purification agent comprises an organic solvent.
23.权利要求 22所述的方法, 其中所述病毒灭活介质固定有有机溶剂病毒灭活 剂和有机溶剂钝化剂, 且所述有机溶剂钝化剂和有机溶剂病毒灭活剂的总含 量大于 0.2 mol/cm3、 优选大于 0.3 mol/cm3The method according to claim 22, wherein the virus inactivating medium is immobilized with an organic solvent virus inactivating agent and an organic solvent deactivator, and the total content of the organic solvent deactivator and the organic solvent virus inactivating agent is More than 0.2 mol/cm 3 , preferably more than 0.3 mol/cm 3 .
24.权利要求 1-23之一所述的方法, 其中所述处理系统还含去白细胞固相介质。 24. The method of any of claims 1-23, wherein the treatment system further comprises a leukocyte-free solid phase medium.
25.权利要求 24所述的方法, 其中所述去白细胞固相介质包括所述病毒灭活剂 吸附介质。 25. The method of claim 24, wherein the leukocyte-free solid phase medium comprises the virus inactivating agent adsorption medium.
26.权利要求 1-25之一所述的方法, 其中所述生物液体包括单人份血液组分。 The method of any one of claims 1 to 25, wherein the biological fluid comprises a single human blood component.
27.权利要求 26所述的方法, 其中所述有机溶剂病毒灭活介质中有机溶剂病毒 灭活剂的含量,等于或小于 3 X (lOppmX所述单人份血液组分的体积 ml)、或 小于(lOppmX所述单人份血液组分的体积 ml)。 The method according to claim 26, wherein the content of the organic solvent virus inactivating agent in the organic solvent virus inactivating medium is equal to or less than 3 X (10 ppm of the volume of the single-part blood component ml), or Less than (lOppmX the volume of the single-part blood component ml).
28.一种有效减小病毒危害的处理系统,其为权利要求 1-27之一所述病毒灭活方 法中所述的处理系统。  28. A treatment system effective to reduce the risk of a virus, which is the treatment system described in the virus inactivation method of any one of claims 1-27.
29.一种有效减小病毒危害的装置, 其至少含权利要求 28所述的处理系统。 29. Apparatus for effectively reducing the risk of a virus, comprising at least the processing system of claim 28.
30.权利要求 29所述的装置, 其为单人份血液组分处理试剂盒。 30. The device of claim 29 which is a single human blood component treatment kit.
PCT/CN2006/000877 2005-04-29 2006-04-29 A method of inactivating virus, treatment system and device used thereof WO2006122476A1 (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110420480A (en) * 2019-08-13 2019-11-08 包头稀土研究院 From the rare earth method for removing alkyl benzene sulphonate ammonium in carbonate precipitating reagent
CN116590245A (en) * 2023-05-18 2023-08-15 吉林和元生物工程股份有限公司 African swine fever virus inactivating agent, virus inactivating method and inactivated virus vaccine

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4540573A (en) * 1983-07-14 1985-09-10 New York Blood Center, Inc. Undenatured virus-free biologically active protein derivatives
US6159375A (en) * 1994-01-10 2000-12-12 Hemasure, Inc. Method for removing leukocytes and methylene blue from plasma
US6348309B1 (en) * 1989-09-13 2002-02-19 Blutspendedienst Der Landesverbaende Des Deutschen Roten Kreuzes Niedersachsen, Oldenburg Und Bremen G.G.M.B.H. Process for inactivating viruses in blood and blood products
CN2595373Y (en) * 2003-01-04 2003-12-31 淄博中保康医疗器具有限公司 Disposable filter for virus inactivation and blood transfusion

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4540573A (en) * 1983-07-14 1985-09-10 New York Blood Center, Inc. Undenatured virus-free biologically active protein derivatives
US6348309B1 (en) * 1989-09-13 2002-02-19 Blutspendedienst Der Landesverbaende Des Deutschen Roten Kreuzes Niedersachsen, Oldenburg Und Bremen G.G.M.B.H. Process for inactivating viruses in blood and blood products
US6159375A (en) * 1994-01-10 2000-12-12 Hemasure, Inc. Method for removing leukocytes and methylene blue from plasma
CN2595373Y (en) * 2003-01-04 2003-12-31 淄博中保康医疗器具有限公司 Disposable filter for virus inactivation and blood transfusion

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110420480A (en) * 2019-08-13 2019-11-08 包头稀土研究院 From the rare earth method for removing alkyl benzene sulphonate ammonium in carbonate precipitating reagent
CN116590245A (en) * 2023-05-18 2023-08-15 吉林和元生物工程股份有限公司 African swine fever virus inactivating agent, virus inactivating method and inactivated virus vaccine
CN116590245B (en) * 2023-05-18 2024-02-09 吉林和元生物工程股份有限公司 African swine fever virus inactivating agent, virus inactivating method and inactivated virus vaccine

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