CN1112937C - Blood plasma virus deactivating method and apparatus - Google Patents

Blood plasma virus deactivating method and apparatus Download PDF

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CN1112937C
CN1112937C CN 98121328 CN98121328A CN1112937C CN 1112937 C CN1112937 C CN 1112937C CN 98121328 CN98121328 CN 98121328 CN 98121328 A CN98121328 A CN 98121328A CN 1112937 C CN1112937 C CN 1112937C
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plasma
photosensitizer
conduit
bag
connected
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CN1249952A (en )
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许亚勇
黄宇闻
张钦辉
钱开诚
高峰
谢如锋
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上海市血液中心
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Abstract

本发明涉及一种血浆病毒的灭活方法及其实施该方法的装置,包括从全血中分离得血浆后,添加光敏剂;以30,000-45,000Lx的荧光强度照射20-80分钟;过滤去除白细胞和光敏剂,这样得到了灭活了病毒的血浆。 After the apparatus relates to a method of plasma viral inactivation methods and embodiments of the present invention, it comprises separating plasma from whole blood obtained was added photosensitizer; irradiation intensity of fluorescence 30,000-45,000Lx 20-80 min; leukocyte-removing filter and photosensitizers, so the virus has been inactivated plasma. 该血浆的整个制备过程在一次性使用、密闭的血液成分采集、分离、去白细胞、吸附光敏剂系统内进行,不需要无菌操作环境;操作方便,安全可靠,易推广应用。 The plasma throughout the preparation of single-use, closed blood component collection, separation, leukocyte, the photosensitizer adsorption system, does not require aseptic environment; easy to operate, reliable, and easy application.

Description

血浆病毒灭活的方法及其装置 Plasma viral inactivation method and apparatus

(1)技术领域本发明涉及临床输注用血浆病毒灭活的方法及实施该方法的装置,特别涉及一种采用光化学方法和过滤、吸附物理方法相结合的血浆病毒的灭活方法及实施该方法的装置。 (1) Technical Field The present invention relates to viral inactivation of plasma in clinical infusion apparatus embodiments of the method and, more particularly to a method and a photochemical filtration, the inactivation method and physical adsorption method embodiment combining plasma virus means method.

(2)背景技术本发明提及的血浆是一种血液成分,它由供血者捐献的全血中分离或用血液单采技术从捐献者体内分离而得,是临床输血治疗一主要的血液成分。 (2) plasma-mentioned background of the invention is a blood component, whole blood from blood donors donate it is separated or isolated from donors with blood apheresis obtained in vivo techniques, is a major clinical transfusion of blood components . 用于临床输注的血浆含有大量不稳定的蛋白、凝血因子,它们往往在制备血浆过程中因环境温度过高而丧失功能。 Plasma for clinical infusion containing a large amount of labile proteins, coagulation factors, they tend to plasma during the preparation of the ambient temperature is too high due to loss of function. 常规制备血浆必须在一定的环境温度下进行,并在血液采集或单采后6小时内完成,置于-20~-40℃冰冻保存。 Conventional plasma must be prepared at a certain temperature, and is completed in 6 hours after blood collection or apheresis, placed in -20 ~ -40 ℃ stored frozen.

光化学灭活病毒的研究始于30年代,自80年代起人们在血浆中加入光敏物质-亚甲蓝,发现其吸收光能后对血浆中的脂包膜和一些非脂包膜病毒具有杀灭作用。 Photochemical Inactivation of the virus began in the 1930s, 80 years since the people to join the photosensitive material in the plasma - methylene blue, found that the absorption of light energy has a killing on plasma lipid-enveloped and non-lipid-enveloped viruses effect. 亚甲蓝灭活病毒的机理是:亚甲蓝与病毒核酸的GC碱基对具有较大的亲和性,当亚甲蓝吸收光能后,可激发产生单态分子氧,这种分子氧的能量形态可破坏病毒的核酸尤其是鸟嘌呤核,从而导致病毒基因的破坏。 Mechanism methylene blue inactivated virus are: methylene blue and GC base pairs of viral nucleic acid has a greater affinity for, when methylene blue absorbs light energy, may produce singlet excited molecular oxygen, molecular oxygen such the form of energy may be destroyed, especially viral nucleic acid is guanine nucleus, resulting in the disruption of the viral genes. 近年来,因输注未经病毒灭活的血浆或血浆制品导致的病毒性传染疾病严重威胁着人们的健康,已经采用的血浆病毒灭活技术如巴斯德法、有机溶剂/表面活性剂法,均因其仅适用于制备血浆制品等大批量样品的处理,而使临床输注的血浆病毒灭活仍处于实验室摸索阶段。 In recent years, due to a viral infection diseases without infusion of viral inactivation of plasma or plasma products cause serious threat to people's health, plasma viral inactivation techniques have been employed, such as Pasteur, organic solvent / surfactant France are its only for processing large numbers of samples were prepared plasma products, the clinical transfusion of plasma virus inactivation is still in the exploratory stage laboratory.

目前国外已有一些厂商针对临床输注血浆的病毒灭活提出了一些实施方案,如美国专利USPat.No.5,639,376A,但他们存在以下问题:1.用于去除白细胞、吸附光敏剂的输血过滤器件没有与血液采集、分离系统组成一体,这在实际操作时血浆易被污染,它对保持被处理血浆中有效成份不受损伤带来诸多不利因素。 At present, some foreign manufacturers for clinical infusion of plasma virus inactivation made some embodiments, such as US Patent USPat.No.5,639,376A, but they are the following problems: 1 for the removal of white blood cells, blood transfusion photosensitizer adsorption filter devices with no blood collection, separation system composed of one, which easily contaminated blood plasma in actual operation, it remains in the plasma treatment active ingredient from damage brought many unfavorable factors.

2.光化学法灭活血浆病毒方法中,所采用的光敏剂剂量虽对人体无毒副作用,但输入人体内不易被排泄,超过一定积累量,仍可能会产生毒副作用。 2. photochemical plasma viral inactivation method, a photosensitizer dose employed although toxic side effects on the human body, but enter the human body can not easily be excreted more than a certain amount of accumulation, it may still produce toxic side effects. 光敏剂的添加量决定病毒灭活的效果,以往的灭活技术忽视了它的添加方式,以及灭活病毒后将其即时排除。 Add the amount of photosensitizer determine the effect of virus inactivation, inactivation technology in the past ignored its way to add, as well as their immediate exclusion will inactivate the virus.

3.给予光敏剂激发态能量的光源可以是全色光,但不同波段不同光强的光源对血浆中蛋白、凝血因子的损伤不一。 3. The light source administering the photosensitizer excited state energy may be full-color light, but the light intensity of the different light sources of different wavelength band in plasma proteins, coagulation factors different damage. 采用荧光照射可最有效地降低对血浆中有效成分损伤。 Using fluorescent light most effectively reduce plasma damage the active ingredient.

目前临床使用的没有光敏剂添加组件和荧光照射塑料袋的四联袋采血密闭系统可进行各种血液成分的制备,其血液成分之一的血浆病毒灭活必须在这一系统中附设光敏剂添加组件和荧光照射塑料袋。 Currently in clinical use and not added fluorescent light component plastic blood collection bags in a closed system quadruple photosensitizer may be prepared by a variety of blood components is performed, one of its plasma viral inactivation of blood components must provide a photosensitizer is added in this system plastic component and fluorescent light. 由该四联袋系统分离的血浆包括单采血浆仍包含约1.0×107的白细胞,这些白细胞携带了大量细胞内病毒,光化学法对其灭活效果较差。 Isolated from the quadruple bag system comprises a plasma leukocyte apheresis plasma still contains from about 1.0 × 107, and a large number of these white blood cells carry the virus, its poor photochemical inactivation method. 另外,光化学法添加的光敏剂浓度虽对人体无毒副作用,但若大量输入人体超出一定的积累量,亦有可能产生毒副作用。 In addition, the concentration of photosensitizer added photochemical method, although toxic side effects on the human body, but if a large number of human input exceeds a certain amount of accumulation, may also produce toxic side effects. 目前虽无明确证据,但其长期滞留于人体的潜在危害不可忽视。 Although there is no clear evidence at present, but its long-term stay in the body's potential hazards can not be ignored.

在临床上很需要没有污染、有效成分不受破坏、已灭活病毒并已去除光敏剂和白细胞的血浆进行输注,以最大限度地降低由于输注血浆对人体造成病毒传染性疾病的传播。 Clinically it needs no pollution, the active ingredient from destruction, the virus has been inactivated and a photosensitizer and white blood cells have been removed infused plasma, in order to minimize the infusion of plasma due to the spread of infectious diseases caused by viruses to humans. 一种理想的、且实际可操作的光化学法灭活临床输注用血浆病毒的流程,应在一密闭的系统内完成。 An ideal, and the actual operational photoinactivation of viruses in clinical plasma infusion process should be completed within a closed system. 这样的系统既可避免灭活病毒操作繁复时间太长影响有效成分失去功能,又可防止操作时可能引起的二次污染以达到既能灭活细胞内、细胞外病毒,又能保证临床输注安全的目的。 Such systems can avoid complicated operations inactivated virus long time lost function affect the active ingredients, may cause secondary pollution can be prevented when another operation to reach both the killed cells, extracellular viruses, but also to ensure clinical infusion security purposes.

(3)发明内容本发明的一个目的是提供既可灭活血浆中游离病毒又能去除血浆中存在的细胞内病毒,集血液采集、血浆分离、去除白细胞、吸附光敏剂为一体的血浆病毒灭活方法。 (3) It is an object of the present invention is to provide a can of the present invention inactivate free virus in plasma can remove the virus present in the plasma cells, blood collection set, plasma separation, the removal of leukocytes, as one of the photosensitizer adsorbed plasma virally live methods.

本发明的再一个目的是提供实施上述方法的装置。 A further object of the present invention is to provide a method of the above-described apparatus embodiment.

本发明的目的是通过下列构思来实现的:一种血浆病毒的灭活方法,其特征在于它包括:从全血中分离得血浆后,以光敏剂在血浆中的最终浓度达0.25-1.5微摩尔/升的量添加光敏剂;以30,000-45,000Lx(勒克斯)的荧光强度照射20-80分钟;过滤去除白细胞和光敏剂,这样得到了灭活了病毒的血浆。 Object of the present invention is achieved by the following concepts: A method for inactivating viruses in plasma, characterized in that it comprises: plasma obtained after the separation, the photosensitizer final concentration of 0.25 to 1.5 in the micro plasma from whole blood mol / liter was added an amount of a photosensitizer; fluorescence irradiation intensity 30,000-45,000Lx (lux) in 20-80 min; leukocyte-removing filter and a photosensitizer, the thus obtained virus inactivated plasma.

其中所述的光敏剂选自亚甲蓝、甲基紫、甲苯胺蓝、局部泌腺540(Merocanine 540,Sigma Chemical Company生产)、补骨脂素(Psoralen,SigmaChemical Company生产)。 Wherein said photosensitizer is selected from methylene blue, methyl violet, toluidine blue, partially glands 540 (Merocanine 540, Sigma Chemical Company, Ltd.), psoralen (Psoralen, SigmaChemical Company, Ltd.).

一种用于灭活血浆病毒的装置,包括光敏剂添加组件,袋体,去白细胞吸附光敏剂输血过滤器件,导管,袋体与光敏剂添加组件、去白细胞吸附光敏剂输血过滤器件组成一密闭系统。 An apparatus for inactivating viruses in plasma, comprising adding a photosensitizer component bag, blood leukocyte adsorption photosensitizer filtration device, a catheter, adding photosensitizer bag body assembly, a photosensitizer adsorbed blood leukocyte filtration device form a closed system.

其中所述的光敏剂添加组件包括易折导通式导管接头、光敏剂加液管和储液管。 Wherein said assembly comprises adding photosensitizers formula frangible guide conduit connector, photosensitizer dosing tube and the reservoir tube.

所述的袋体包括采血袋、转移袋、荧光照射袋和血浆储存袋。 The collecting bag comprises a bag body, transfer bags, bags and fluorescent light plasma storage bag.

(4)附图说明图1是灭活血浆病毒的方法流程图。 (4) FIG. 1 is a flowchart of a method of inactivating viruses in plasma.

图2是实施所述灭活血浆病毒方法的装置的示意图。 FIG 2 is a schematic diagram of the apparatus of plasma viral inactivation method embodiment.

图3是易折导通式光敏剂添加组件的示意图。 3 is a schematic assembly guide formula frangible added photosensitizer.

图4是本发明装置的较佳实施方案。 FIG 4 is a preferred embodiment of the apparatus of the present invention.

图5是温控平行振摇式荧光照射仪控制示意图。 FIG 5 is a temperature-controlled shaking parallel fluorescent light scanner control schematic formula.

(5)具体实施方式下面结合附图对本发明作进一步阐述。 DRAWINGS The invention will be further illustrated (5) below in connection with specific embodiments.

参见附图2,一种实施灭活血浆病毒方法的装置,包括采血针1,采血袋3,导管2、4、5、7、8、10、12、13和16,第一转移袋6,易折导通式光敏剂添加组件9,荧光照射袋11,去白细胞吸附光敏剂输血过滤器件15和血浆储存袋17,所述的采血针1通过导管2与采血袋3连接,一端连接采血袋3的导管4分支成导管5和7,所述的导管5与第一转移袋6连接,所述的导管7与导管8相连,所述的导管8与所述的易折导通式光敏剂添加组件9相连,所述的添加组件9通过导管10与荧光照射袋11相连,一端连接荧光照射袋11的导管12分支成导管13和带有所述去白细胞吸附光敏剂输血过滤器件15的导管,然后所述的两个导管汇集成导管16,通向血浆储存袋17。 Referring to Figure 2, an apparatus for plasma viral inactivation method embodiments, including a lancet, blood collection bag 3, the catheter 2,4,5,7,8,10,12,13 and 16, the first transfer bag 6, adding a photosensitizer formula frangible guide assembly 9, the bag 11 fluorescent light, a photosensitizer adsorbed leukocyte transfusion filtering device 15 and the plasma storage bag 17, the blood collection needle 1 through a conduit connected to the blood collection bag 2 and 3, one end connected to the blood collection bag the conduit 4 branches into three conduits 5 and 7, the first transfer conduit 5 and the bag 6 connected to a conduit 7 connected to the catheter 8, the catheter 8 guiding the frangible photosensitizer formula 9 is connected to add components, the addition of component 9 connected to the conduit 10 and the fluorescent light 11 through the bag, the bag having one end connected fluorescent light conduit 11 is branched into the conduit 12 with the conduit 13 and leukocyte adsorption photosensitizer transfusion filtering device 15 then the two conduits together into the conduit 16 leading to the plasma storage bag 17. 在一个实施方案中,所述的导管13上带有排气夹14。 In one embodiment, the conduit 13 with the exhaust clamp 14.

采血袋3的容积可为常规大小,一般是200毫升,第一转移袋6、荧光照射袋11和血浆储存袋17的容积大小也可由本技术领域人员根据血浆的容量作出常规的选择,较好的是200毫升。 Volume of the blood collection bag 3 may be of conventional size, generally 200 ml, the first transfer bag 6, the bag 11 and the fluorescent light volume of plasma storage bag 17 can also be the size of those in the art to make conventional selection according to the capacity of the plasma, preferably is 200 ml. 所述袋子的材料可为医药上可接受的聚合物材料,如聚氯乙烯等,导管材料可为聚氯乙烯等。 The bag material may be a pharmaceutically acceptable polymer material, such as polyvinyl chloride, the catheter material may be polyvinyl chloride.

去白细胞吸附光敏剂输血过滤器件15包括至少一个进口和一个出口的塑料外壳,用作滤除白细胞的滤芯材料可以是聚丙烯、聚酯不织布,无钙玻璃纤维、醋酸纤维或它们与天然棉纤维的混合物和对光敏剂亚甲蓝及其产物有强吸附作用的活性碳纤维。 Adsorption of photosensitizer leukocyte transfusion filtering device 15 comprises at least one inlet and an outlet of the plastic housing, the filter material was filtered off as white blood cells may be polypropylene, polyester non-woven, non-lime glass, cellulose acetate or mixtures thereof with natural cotton fibers and mixtures of strong adsorption of Methylene Blue and the product activated carbon fiber. 通过改变这些滤芯材料表面特征,与血浆的接触截面积或滤芯的填充量,调节自重流动速率,实现白细胞去除率大于99.9%。 By changing the filter characteristics of these surface materials, the amount of plasma or fill the contact cross-sectional area of ​​the filter, adjusting the weight flow rate, to achieve greater than 99.9% removal of leukocytes.

进一步的是,过滤部件中还含有活性碳纤维,它对光敏剂及其产物有强的吸附作用,在去除白细胞的同时将85%以上的光敏剂吸附,使其对人体的影响降到最低限度。 Further, the filter member further comprising activated carbon fiber, and its product photosensitizers strong adsorption, while removing leukocytes will be more than 85% adsorption of the photosensitizer, its impact on the human body to a minimum.

参见图3,所述的易折导通式光敏剂添加组件9包括易折导通式导管接头20,光敏剂加液管21,储液管22和易折封闭头23。 Referring to Figure 3, the frangible guide assembly 9 comprises adding the photosensitizer formula frangible Formula guide conduit connector 20, the photosensitizer dosing tube 21, reservoir tube 22 is closed off head 23 is easy. 用于光敏剂添加组件的材料可为医用高分子塑料避光材料,如含有色母的聚氯乙烯或医用PBS。 Adding material for the photosensitizer medical polymer component may be a dark plastic material, such as polyvinyl chloride or medical masterbatch containing PBS.

其中储液管22的容积可为3-5毫升,可由含有棕色色母的聚氯乙烯制成,易折封闭头23可由硬性聚氯乙烯注塑制成,这样只需用手一折,封闭头23即被打开,从而使导管8与组件9、组件9与导管10得以沟通。 Wherein the volume of the reservoir tube 22 may be 3-5 ml, may be made of polyvinyl chloride containing a brown masterbatch made frangible closure head 23 may be rigid PVC injection molding, so that only a hand-off, the closing head 23 i.e. open, so that the catheter assembly 8 and 9, the catheter assembly 10 9 to communicate.

参见图4,在本发明的较佳技术方案中,所述的灭活病毒装置可进一步包括导管18和血小板保存袋19。 Referring to Figure 4, in a preferred aspect of the present invention, the inactivated virus may further comprise conduit means 18 and the platelet preserving bag 19. 在离心后,采血袋3中放置的全血被分离成血浆和血小板,所述的血浆经导管4和7进入荧光照射袋11,所述的血小板经导管7和18进入血小板保存袋19供临床上的使用。 After centrifugation, whole blood collection bag 3 placed in blood is separated into plasma and platelets, the plasma via conduit 7 enters the fluorescent light 4 and the bag 11, the platelet 18 via conduit 7 and into the platelet preserving bag 19 for clinical use on.

将用采血针1收集的全血放在采血袋3中,离心分离后获得的血浆经导管4和5放在第一转移袋6中,折断易折导通式导管接头20中的易折封闭头23,使血浆经导管7、8与易折导通式光敏剂添加组件9内的光敏剂混合,其中这样选择光敏剂的量,使光敏剂在血浆中的最终浓度为0.25-1.5微摩尔/升,然后血浆和光敏剂混合经导管10进入荧光照射塑料袋11,高频热合导管10。 The blood bag 3 on the whole blood collected by the blood collection needle 1, the plasma obtained after centrifugation through conduit 4 and 5 in the first transfer bag 6, breaking frangible guide conduit connector 20 in the general formula frangible closure head 23, the plasma via conduit 7, 8 and the guide frangible formula photosensitizers added in the mixing assembly 9 photosensitizer, wherein the amount of the photosensitizer is selected such that the final concentration of the photosensitizer in the plasma was 0.25 and 1.5 micromolar / liter, and mixing the plasma and a photosensitizer via conduit 10 into the bag 11 fluorescent light, high-frequency heat pipe 10. 将含有血浆的荧光照射塑料袋11、去白细胞吸附光敏剂输血过滤器件15、血浆储存袋17等组件,置于温控平行振摇式荧光照射仪中,根据图5所示的温控平行振摇式荧光照射仪控制示意图的工作方式,在4-18℃下于光强度为30,000-45,000Lx下照射20-80分钟来灭活病毒,然后在去白细胞吸附光敏剂输血过滤器件15以自重流动工作方式、过滤速度为10-35毫升/分钟来去除由白细胞携带的病毒并吸附光敏剂,再经导管16流入储存袋17,在冷冻保存前用排气夹14排去空气,高频热合导管16,将血浆储存袋17保存在-20~-40℃环境下供临床输注或制备血浆制品用。 The fluorescent light plastic bag 11 containing the plasma, adsorption of leukocytes to the photosensitizer transfusion filtering device 15, plasma components such as storage bags 17, disposed in parallel with shaking thermostat formula fluorescent light meter, thermostat parallel vibration as shown in FIG. 5 rocking schematic fluorescent light scanner control work, at 4-18 deg.] C in the irradiation light intensity for the next 20-80 minutes 30,000-45,000Lx to inactivate viruses, then filtered blood leukocyte adsorption photosensitizer flow device 15 by its own weight mode of operation, the filtration rate of 10-35 ml / min to remove the virus and adsorption of leukocytes carried photosensitizer, and then flows into the storage bag 16 via a conduit 17, prior to cryopreservation air exhaust 14 to the exhaust clip, high-frequency heat pipe 16, the plasma storage bag 17 for clinical infusions stored at -20 ~ -40 ℃ environment or plasma products prepared with.

实施例110℃的环境温度下,在全血经离心后,分离得到123毫升血浆。 At ambient temperature 110 ℃ embodiment embodiment, the whole blood by centrifugation, and 123 ml of plasma separation. 分离后得到的血小板经导管7到导管18进入血小板保存袋19。 Platelets obtained after separation via conduit 7 to the conduit 18 into the platelet preserving bag 19. 血浆经导管4、7、8、折断易折导通式导管接头20中的易折封闭头23与易折导通式光敏剂添加组件9内的0.123微摩尔亚甲蓝(泰兴市制药厂生产)混合,然后血浆和光敏剂混合物经导管10进入荧光照射袋11。 Plasma catheter 4,7,8, breaking frangible guide catheter formula frangible joint 20 closing head 23 and the frangible component is added photosensitizer guide Formula 9 0.123 micromolar methylene blue (Taixing pharmaceutical production ), and then into the plasma, and a mixture of fluorescent light photosensitizer bags 11 via conduit 10. 将含有血浆的荧光照射塑料袋11、去白细胞吸附光敏剂输血过滤器件15、血浆储存袋17等组件,置于温控平行振摇式荧光照射仪中,在4℃下于光强度为38,000Lx下照射30分钟来灭活病毒,然后在过滤、吸附截面积为12厘米2的去白细胞吸附光敏剂输血过滤器件15中以自重流动工作方式、过滤速度为25-35毫升/分钟来去除由白细胞携带的病毒并吸附亚甲蓝,再经导管16流入储存袋17。 The fluorescent light plastic bag 11 containing the plasma, adsorption of leukocytes to the photosensitizer transfusion filtering device 15, plasma components such as storage bags 17, disposed in parallel with shaking thermostat formula fluorescent light instrument, the light at an intensity of 4 ℃ 38,000Lx irradiated for 30 minutes to inactivate the virus, and then filtered, the adsorption cross sectional area of ​​12 cm 2 photosensitizer adsorbed blood leukocyte filter device 15 to the weight flow mode of operation, the filtration rate of 25-35 ml / min to remove the leukocytes carrying virus and adsorption of methylene blue, and then flows into the storage bag 16 via conduit 17. 用VSV、Sindbis指示病毒测量血浆logTCID50的变化,结果如下表1所示。 With VSV, Sindbis virus indicating changes in plasma logTCID50 measurement results are shown in Table 1. 在冷冻保存前用排气夹14排去空气,将血浆储存袋17保存在-20~-40℃环境下供临床输注或制备血浆制品用,血小板保存袋19置于22℃环境备用。 Before cryopreservation exhaust clamp 14 to exhaust air, the plasma storage bag 17 for clinical infusions stored at -20 ~ -40 ℃ environment or article prepared by plasma, the platelet preserving bag 19 placed in 22 ℃ RESERVE.

根据实施例1所述的方法,除了荧光照射的时间30分钟被照射时间5分钟、10分钟和15分钟替代外其它基本根据实施例1所述的方法得到了血浆制品,用VSV,Sindbis指示病毒测定荧光照射时间不同对灭活血浆病毒效果的影响,结果如下表1所示。 The method of Example 1, except time of fluorescent light for 30 minutes irradiation time was 5 minutes, 10 minutes and 15 minutes have been replaced by other blood products in accordance with the basic embodiment of the method of Example 1, indicating a virus with VSV, Sindbis fluorescence was measured on the inactivation of different irradiation time of plasma viral effects, the results shown in table 1 below.

表 1亚甲蓝+荧光照射不同时间含VSV,Sindbis病毒血浆log TCID50的变化光照时间(分钟)病毒 滴度 100ml血浆稀释 0(含MB) 5 10 15 30VSV 8.5 6.38 4.83 1.5 1.5 0.5 0.5Sindbis 8.5 6.33 4.5 2.33 1.25 0.5 0.5按照实施例1所述的方法,分别对下表2所列出的血浆进行灭活病毒处理,结果如下表2所示。 Table 1 + methylene blue fluorescent light at different times containing VSV, Sindbis virus illumination time change log TCID50 plasma (min) virus titer plasma was diluted 100ml 0 (including MB) 5 10 15 30VSV 8.5 6.38 4.83 1.5 1.5 0.5 0.5Sindbis 8.5 6.33 2.33 1.25 4.5 0.5 0.5 the method described in Example 1, the plasma are listed in table 2 virus inactivation treatment, the results shown in table 2 below.

表2 亚甲蓝/荧光照射煤火血浆病毒实施例结果序 血浆 亚甲 剩余 血浆蛋白回收率 凝血因子回收率号 体积 蓝去 白细除率 胞(mL) (%) (×104IgG IgM IgA Fg II VII VIII IX PLG AT-III)1 123 ≥85.0 ≤1.0 87.30 92.77 100. 81.70 77.78 84.71 86.67 100. 98.59 75.862 117 ≥85.0 ≤1.0 83.56 100. 79.67 82.74 82.02 100. 71.21 81.2 75.32 90.1253 109 ≥85.0 ≤1.0 100. 81.21 100. 87.82 78.65 100. 100. 71.2 98.21 98.6514 132 ≥80.0 ≤1.0 95.81 100. 95.12 100. 94.25 100. 95.81 100. 86.57 94.125 105 ≥90.0 ≤1.0 90.91 99.00 100. 92.58 98.84 76.90 90.91 91.3 67.02 10006 116 ≥85.0 ≤1.0 98.08 88.68 94.79 73.99 78.65 87.38 98.08 73.1 85.53 90.2837 106 ≥85.0 ≤1.0 100. 100. 100. 100. 100. 91.96 100. 90.7 73.17 95.4548 103 ≥90.0 ≤1.0 88.69 Cell rate (mL) (%) (× 104IgG IgM IgA Fg II in Table 2 except methylene blue / fluorescent light coals results of Examples plasma viral sequence methylenedioxy plasma coagulation factor recovery of residual plasma protein recovery volume number blue to white fine VII VIII IX PLG AT-III) 1 123 ≥85.0 ≤1.0 87.30 92.77 100. 81.70 77.78 84.71 86.67 100. 98.59 75.862 117 ≥85.0 ≤1.0 83.56 100. 79.67 82.74 82.02 100. 71.21 81.2 75.32 90.1253 109 ≥85.0 ≤1.0 100. 81.21 100. 87.82 78.65 100. 100. 71.2 98.21 98.6514 132 ≥80.0 ≤1.0 95.81 100. 95.12 100. 94.25 100. 95.81 100. 86.57 94.125 105 ≥90.0 ≤1.0 90.91 99.00 100. 92.58 98.84 76.90 90.91 91.3 67.02 10006 116 ≥85.0 ≤1.0 98.08 88.68 94.79 73.99 78.65 87.38 98.08 73.1 85.53 90.2837 106 ≥85.0 ≤1.0 100. 100. 100. 100. 100. 91.96 100. 90.7 73.17 95.4548 103 ≥90.0 ≤1.0 88.69 96.84 84.27 82.74 86.42 91.67 88.69 89.8 91.84 100. 96.84 84.27 82.74 86.42 91.67 88.69 89.8 91.84 100.

39 121 ≥85.0 ≤1.0 91.67 100. 100. 100. 82.72 89.32 91.67 100. 100. 100.10 107 ≥85.0 ≤1.0 100. 90.00 100. 72.67 100. 71.43 100. 100. 76.19 93.65本发明采用光化学,并结合物理方法,制备经病毒灭活、去白细胞的新鲜冰冻血浆,可灭活细胞内、细胞外的病毒。 39 121 ≥85.0 ≤1.0 91.67 100. 100. 100. 82.72 89.32 91.67 100. 100. 100.10 107 ≥85.0 ≤1.0 100. 90.00 100. 72.67 100. 71.43 100. 100. 76.19 93.65 photochemical the present invention, a combination of physical methods and the preparation virally inactivated, fresh frozen plasma to leukocytes, inactivated cells, extracellular virus. 整个制备过程在一次性使用、密闭的血液成分采集、分离、去白细胞、吸附光敏剂系统内进行,不需要无菌操作环境;病毒滴度下降6logTCID50以上,剩余白细胞数小于1.0×104,光敏剂的去除率大于85%,凝血因子回收率大于80%;操作方便,安全可靠,易推广应用。 Throughout the preparation of single-use, closed blood component collection, separation, leukocyte, the photosensitizer adsorption system, does not require aseptic environment; viral titer decreased 6logTCID50 above, the residual leukocyte count is less than 1.0 × 104, photosensitizer removal efficiency greater than 85%, greater than 80% recovery of clotting factor; easy to operate, reliable, and easy application. 所述经灭活病毒的血浆在常规的冰冻保存条件下能保存2年,大大方便了临床上的使用。 The virus inactivated plasma stored frozen under conventional conditions can be stored for 2 years, greatly facilitate the clinical use.

Claims (6)

  1. 1.一种血浆病毒的灭活方法,其特征在于它包括:从全血中分离得血浆后,以光敏剂在血浆中的最终浓度达0.25-1.5微摩尔/升的量添加选自亚甲蓝、甲基紫、甲苯胺蓝、局部泌腺540或补骨脂素的光敏剂;以30,000-45,000勒克斯的荧光强度照射20-80分钟;过滤去除白细胞和所述的光敏剂,这样得到了灭活了病毒的血浆。 CLAIMS 1. A method of inactivating viruses in plasma, characterized in that it comprises: plasma obtained after the separation, the photosensitizer final concentration in plasma from the whole blood of 0.25 to 1.5 micromoles / liter was added an amount selected from methylene blue, methyl violet, toluidine blue, partially glands psoralen photosensitizers or 540; 30,000 to 45,000 lux at an irradiation intensity of fluorescence 20-80 min; filtered to remove leukocytes and the photosensitizer, the thus obtained inactivated plasma virus.
  2. 2.根据权利要求1所述的灭活方法,其特征在于所添加的光敏剂在血浆中的最终浓度为1.0微摩尔/升。 2. The method of inactivation according to claim 1, characterized in that the final concentration of the photosensitizer in the added plasma was 1.0 mol / liter.
  3. 3.一种用于灭活血浆病毒的装置,其特征在于它包括采血针(1),采血袋(3),导管(2)、(4)、(5)、(7)、(8)、(10)、(12)、(13)和(16),第一转移袋(6),易折导通式光敏剂添加组件(9),荧光照射袋(11),去白细胞吸附光敏剂输血过滤器件(15)和血浆储存袋(17),所述的采血针(1)通过导管(2)与采血袋(3)连接,一端连接采血袋(3)的导管(4)分支成导管(5)和(7),所述的导管(5)与第一转移袋(6)连接,所述的导管(7)与导管(8)相连,所述的导管(8)与所述的易折导通式光敏剂添加组件(9)相连,所述的添加组件(9)通过导管(10)与荧光照射袋(11)相连,一端连接荧光照射袋(11)的导管(12)分支成导管(13)和带有所述去白细胞吸附光敏剂输血过滤器件(15)的导管,然后所述的两个导管汇集成导管(16),通向血浆储存袋(17)。 An apparatus for inactivating viruses in plasma, characterized in that it comprises a lancet (1), blood bags (3), the catheter (2), (4), (5), (7), (8) , (10), (12), (13) and (16), a first transfer bag (6), adding a photosensitizer formula frangible guide assembly (9), fluorescent light bags (11), adsorption of leukocytes to the photosensitizer transfusion filtering device (15) and the plasma storage bag (17), said lancet (1) connected by a conduit (2) and the blood collection bag (3) having one end connected to the blood collection bag (3) of the conduit (4) branches into conduit (5) and (7), said conduit (5) connected to the first transfer bag (6), said duct (7) is connected to the conduit (8), said conduit (8) with said formula photosensitizer added frangible guide assembly (9), said adding component (9) connected via conduit (10) and the fluorescent light bag (11) having one end connected to the bag fluorescent light (11) of the conduit (12) branches into the conduit (13) and with said suction conduit photosensitizer leukoreduced transfusion filtering device (15), and said two conduits together into the conduit (16) leading to the plasma reservoir bag (17).
  4. 4.根据权利要求3所述的装置,其特征在于,所述的导管(13)上带有排气夹(14)。 4. The apparatus according to claim 3, characterized in that said exhaust duct with clamp (14) (13).
  5. 5.根据权利要求4所述的装置,其特征在于所述的易折导通式光敏剂添加组件(9)包括易折导通式导管接头(20),光敏剂加液管(21),储液管(22)和易折封闭头(23)。 5. The apparatus as claimed in claim 4, wherein said photosensitizer is added frangible guide assembly Formula (9) comprises a frangible linker guide catheter formula (20), a photosensitizer plus liquid pipe (21), a reservoir tube (22) and ease of folding of the closing head (23).
  6. 6.根据权利要求3,4或5所述的装置,其特征在于该装置进一步包括血小板保存袋(19),它通过与所述导管(7)相连的导管(18)与所述的装置相连。 6. The apparatus of claim 4 or claim 5, characterized in that the apparatus further comprising a platelet preserving bag (19) which is connected to the device via conduit (18) connected to said conduit (7) .
CN 98121328 1998-10-07 1998-10-07 Blood plasma virus deactivating method and apparatus CN1112937C (en)

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US7445756B2 (en) 1999-06-03 2008-11-04 Fenwal, Inc. Fluid processing sets and organizers for the same
EP1555027A4 (en) 2002-10-16 2008-06-04 Asahi Kasei Medical Co Ltd Plasma preparation or serum preparation and process for producing the same
US8808977B2 (en) 2003-05-22 2014-08-19 Beijing Jingjing Medical Equipment Co., Ltd Method of inactivating virus in circular blood and its applications in treating viral diseases
CN1317038C (en) * 2003-07-03 2007-05-23 北京京精医疗设备有限公司 Virus inactivating method of extracorporeal circulated blood
CN100413542C (en) 2006-06-01 2008-08-27 上海市血液中心;南京逐陆医药科技有限公司 Pathogen inactivator composition for blood or blood component and its compounding process
CN100417722C (en) 2006-10-25 2008-09-10 中国辐射防护研究院 Inactivating method for virus in blood products
CN103861132A (en) * 2012-12-18 2014-06-18 苏州排头兵药业科技有限公司 Plasma and plasma product virus inactivation device
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