CN1249952A - Blood plasma virus deactivating method and apparatus - Google Patents
Blood plasma virus deactivating method and apparatus Download PDFInfo
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- CN1249952A CN1249952A CN 98121328 CN98121328A CN1249952A CN 1249952 A CN1249952 A CN 1249952A CN 98121328 CN98121328 CN 98121328 CN 98121328 A CN98121328 A CN 98121328A CN 1249952 A CN1249952 A CN 1249952A
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Abstract
The deactivating process including separation of whole blood to obtain blood plasma, addition of photosensitizer, irradiation with fluorescent light of 30,000-45,000 Lx intensity for 20-80 min and filtration to eliminate leucocyte and photosensitizer. The whole process is completed in an disposable sealed system for blood collection, separation and elimination of leucocyte and photosensitizer and needs no bacteria-free operation environment. The method and apparatus are convenient, safe, reliable and applicable.
Description
The present invention relates to clinical infusion with the method for blood-plasma virus killing and implement the device of this method, particularly a kind of device that adopts the ablation method of the plasma viral that photochemical method and filtration, absorption physical method combine and implement this method.
The blood plasma that the present invention mentions is a kind of blood constituent, separates in the whole blood that it is contributed by the blood donor or separates in donor's body and get with the blood list technology of adopting, and be that clinical blood transfusion is treated a main blood constituent.The blood plasma that is used for clinical infusion contains a large amount of unsettled albumen, thrombin, they often in preparation blood plasma process because of the too high loss of function of ambient temperature.Conventional preparation blood plasma must carry out under certain ambient temperature, and at blood collection or singly adopt in back 6 hours and finish, places-20～-40 ℃ of stored frozen.
The research of photochemistry inactivation of viruses starts from the thirties, and people add photoactive substance-methylene blue in blood plasma from the eighties, finds after it absorbs luminous energy the fat peplos in the blood plasma and some non-lipid-coated virus to be had killing action.The mechanism of methylene blue inactivation of viruses is: the G-C base pair of methylene blue and viral nucleic acid has bigger affinity, behind methylene blue absorption luminous energy, can excite and produce the singlet molecular oxygen, but the nucleic acid of the energy form break virus of this molecular oxygen is guanosint especially, thereby the destruction that causes viral gene.In recent years, because of infusion people's health without the viral communicate illness serious threat that the blood plasma or the blood plasma product of inactivation of virus causes, blood-plasma virus killing technology that has adopted such as pasteur's method, organic solvent/surfactant method, all be only applicable to prepare the processing of batch samples such as blood plasma product, and make the blood-plasma virus killing of clinical infusion still be in the laboratory stage of fumbling because of it.
External at present more existing manufacturers have proposed some embodiments at the inactivation of virus of clinical infusion blood plasma, as United States Patent (USP) U.S.Pat.No.5, and 639,376A, but there is following problem in they:
1. the transfusion filter spare that is used to remove leukocyte, absorption photosensitizer not and blood collection, piece-rate system form one, this blood plasma when practical operation is easily contaminated, active ingredient is injury-free to bring many unfavorable factors to keeping in the processed blood plasma for it.
2. in the photochemical method deactivation plasma viral method,, be difficult for being drained in the input human body, surpass certain accumulation, still may produce toxic and side effects though the photosensitizer dosage that is adopted has no side effect to human body.The effect of the addition of photosensitizer decision inactivation of virus, inactivation technology has in the past been ignored its addition manner, and behind the inactivation of viruses with its instant eliminating.
3. the light source that gives the photosensitizer excited energy can be panchromatic light, but the light source of different-waveband different light intensity differs to the damage of albumen, thrombin in the blood plasma.Adopt the fluorescence irradiation can reduce most effectively to effective ingredient damage in the blood plasma.
The tetrad bag blood sampling closed system that does not have photosensitizer to add assembly and fluorescence irradiated plastics bag of present clinical use can carry out the preparation of various blood constituents, and the blood-plasma virus killing of one of its blood constituent must be set up photosensitizer and add assembly and fluorescence irradiated plastics bag in this system.Comprise that by the isolating blood plasma of this tetrad bag system Plasma Pheresis/Apheresis Plasma still comprises about 1.0 * 10
7Leukocyte, these leukocyte have carried a large amount of intracellular virus, photochemical method is relatively poor to its inactivating efficacy.In addition, though the photosensitizer concentration that photochemical method adds has no side effect to human body,, also might produce toxic and side effects if a large amount of input human body exceeds certain accumulation.Though do not have clear and definite evidence at present, its potential hazard that is stranded in human body for a long time is very important.
Be sought after clinically not polluting, effective ingredient is not damaged, inactivation of viruses and removed photosensitizer and leukocytic blood plasma carries out infusion, to reduce to greatest extent because infusion blood plasma causes the propagation of viral infectious disease to human body.The clinical infusion of a kind of ideal and actual exercisable photochemical method deactivation is with the flow process of plasma viral, should finish in an airtight system.Such system both can avoid inactivation of viruses to operate the oversize effective ingredient that influences of complicated time and lose function, the secondary pollution that may cause in the time of can preventing to operate again with reach can as killed cells in, extracellular virus, can guarantee clinical infusion purpose of safety again.
But an object of the present invention is to provide not only in the deactivation blood plasma free virus but also can remove the intracellular virus that exists in the blood plasma, collection blood collection, separating plasma, removal leukocyte, absorption photosensitizer are the blood-plasma virus killing method of one.
A further object of the present invention provides the device of implementing said method.
The objective of the invention is to realize by following design:
A kind of ablation method of plasma viral is characterized in that it comprises: from whole blood, separate behind the blood plasma, the amount that reaches the 0.25-1.5 micromoles per liter with the ultimate density of photosensitizer in blood plasma is added photosensitizer; With 30,000-45, the fluorescence intensity irradiation of 000Lx 20-80 minute; Filtering removing leukocytes and photosensitizer, the viral blood plasma that obtained deactivation like this.
Wherein said photosensitizer is selected from methylene blue, crystal violet, toluidine blue, part and secretes gland 540 (Merocanine540, Sigma Chemical Company produces), psoralen (Psoralen, Sigma Chemical Company produces).
A kind of device that is used for the deactivation plasma viral comprises that photosensitizer adds assembly, and bag, leucocyte-removing absorption photosensitizer transfusion filter spare, conduit, bag and photosensitizer add assembly, leucocyte-removing absorption photosensitizer transfusion filter spare is formed a closed system.
Wherein said photosensitizer adds assembly and comprises conduction type catheter adapter easy to break, photosensitizer liquid-feeding tube and liquid storage pipe.
Described bag comprises blood taking bag, transfering bag, fluorescence irradiation bag and blood plasma reservoir bag.
Fig. 1 is the method flow diagram of deactivation plasma viral.
Fig. 2 is a schematic representation of apparatus of implementing described deactivation plasma viral method.
Fig. 3 is the sketch map that conduction type photosensitizer easy to break adds assembly.
Fig. 4 is the preferable embodiment of apparatus of the present invention.
Fig. 5 is the parallel jolting formula of a temperature control fluorescence radiation instrument control sketch map.
The present invention is further elaborated below in conjunction with accompanying drawing.
Referring to accompanying drawing 2, a kind of device of implementing deactivation plasma viral method, comprise blood taking needle 1, blood taking bag 3, conduit 2,4,5,7,8,10,12,13 and 16, the first transfering bags 6, conduction type photosensitizer easy to break adds assembly 9, fluorescence irradiation bag 11, leucocyte-removing absorption photosensitizer transfusion filter spare 15 and blood plasma reservoir bag 17.
The volume of blood taking bag 3 can be conventional size, generally is 200 milliliters, and the volume size of first transfering bag 6, fluorescence irradiation bag 11 and blood plasma reservoir bag 17 also can be made conventional selection according to the capacity of blood plasma by present technique field personnel, is preferably 200 milliliters.The material of described sack can be pharmaceutically acceptable polymeric material, and as polrvinyl chloride etc., tube material can be polrvinyl chloride etc.
Leucocyte-removing absorption photosensitizer transfusion filter spare 15 comprises the cabinet of at least one import and an outlet, as the leukocytic material for core of filtering can be polypropylene, polyester adhesive-bonded fabric, the mixture of no lime glass fiber, acetate fiber or they and natural cotton fiber and photosensitizer methylene blue and product thereof had strong adsorbing activated carbon fiber.By changing these material for core surface character, the loading of or filter element long-pending with the contacting section of blood plasma is regulated the deadweight flow rate, realizes that leukocyte removal efficiency is greater than 99.9%.
Further be, also contain activated carbon fiber in the filter element, it has strong adsorption to photosensitizer and product thereof, and is leukocytic simultaneously with the absorption of the photosensitizer more than 85% in removal, makes its influence to human body drop to bottom line.
Referring to Fig. 3, described conduction type photosensitizer easy to break adds assembly 9 and comprises conduction type catheter adapter 20 easy to break, photosensitizer liquid-feeding tube 21, liquid storage pipe 22 and closed head easy to break 23.The material that is used for photosensitizer interpolation assembly can be medical high polymer plastics lucifuge material, as contains coloured mother's polrvinyl chloride or medical PBS.
Wherein the volume of liquid storage pipe 22 can be the 3-5 milliliter, can be made by the polrvinyl chloride that contains brown color mother, closed head 23 easy to break can be made by the rigid polyvinyl chloride injection moulding, like this only with hand one folding, closed head 23 promptly is opened, thereby conduit 8 and assembly 9, assembly 9 are linked up with conduit 10.
Referring to Fig. 4, in preferred technique scheme of the present invention, described inactivation of viruses device can further comprise conduit 18 and blood platelet storage bag 19.After centrifugal, the whole blood of placing in the blood taking bag 3 is separated into blood plasma and platelet, and described blood plasma enters fluorescence irradiation bag 11 through conduit 4 and 7, and described platelet enters blood platelet storage bag 19 for clinically use through conduit 7 and 18.
To be placed in the blood taking bag 3 with the whole blood that blood taking needle 1 is collected, the blood plasma that obtains after the centrifugalize is placed in first transfering bag 6 through conduit 4 and 5, closed head easy to break 23 in the conduction type catheter adapter 20 easy to break fractures, blood plasma is mixed with photosensitizer in conduction type photosensitizer easy to break adds assembly 9 through conduit 7,8, wherein select the amount of photosensitizer like this, making the ultimate density of photosensitizer in blood plasma is the 0.25-1.5 micromoles per liter, the mixing of blood plasma and photosensitizer enters fluorescence irradiated plastics bag 11, high frequency heat seal conduit 10 through conduit 10 then.The fluorescence irradiated plastics bag 11 that will contain blood plasma, leucocyte-removing absorption photosensitizer transfusion filter spare 15, blood plasma reservoir bag 17 assemblies such as grade, place the parallel jolting formula of temperature control fluorescence radiation instrument, working method according to the parallel jolting formula of temperature control shown in Figure 5 fluorescence radiation instrument control sketch map, under 4-18 ℃, be 30 in light intensity, 000-45,000Lx irradiation down came inactivation of viruses in 20-80 minute, adsorb photosensitizer transfusion filter spare 15 with the mobile working method of conducting oneself with dignity at leucocyte-removing then, the rate of filtration removes the virus of being carried by leukocyte and adsorbs photosensitizer for the 10-35 ml/min, flow into reservoir bag 17 through conduit 16 again, before freezing preservation, press from both sides 14 venting air with aerofluxus, high frequency heat seal conduit 16 is kept under-20～-40 ℃ of environment blood plasma reservoir bag 17 for clinical infusion or preparation blood plasma product usefulness.
Under 10 ℃ the ambient temperature, after centrifugal, separate obtaining 123 milliliters of blood plasma at whole blood.The platelet that obtains after the separation enters blood platelet storage bag 19 through conduit 7 to conduit 18.The easy to break closed head 23 of blood plasma in conduit 4,7,8, the conduction type catheter adapter 20 easy to break that fractures mixes with 0.123 micromole's methylene blue (productions of Taixing City pharmaceutical factory) in conduction type photosensitizer easy to break adds assembly 9, and blood plasma and photosensitizer mixture enter fluorescence through conduit 10 and shine bags 11 then.Fluorescence irradiated plastics bag 11, the leucocyte-removing that will contain blood plasma adsorb assemblies such as photosensitizer transfusion filter spare 15, blood plasma reservoir bag 17, place the parallel jolting formula of temperature control fluorescence radiation instrument, under 4 ℃, be 38 in light intensity, 000Lx irradiation down came inactivation of viruses in 30 minutes, was 12 centimetres filtering, adsorbing sectional area then
2Leucocyte-removing absorption photosensitizer transfusion filter spare 15 in be that the 25-35 ml/min removes the virus of being carried by leukocyte and adsorbs methylene blue with the mobile working method of conducting oneself with dignity, the rate of filtration, flow into reservoir bags 17 through conduit 16 again.With the variation that VSV, Sindbis indicator virus are measured blood plasma logTCID50, the result is as shown in table 1 below.Press from both sides 14 venting air with aerofluxus before freezing preservation, blood plasma reservoir bag 17 is kept at supplies clinical infusion or preparation blood plasma product usefulness under-20～-40 ℃ of environment, blood platelet storage bag 19 places 22 ℃ of environment standby.
According to embodiment 1 described method, other has obtained blood plasma product according to embodiment 1 described method substantially except the 5 minutes illuminated time of 30 minutes time, 10 minutes and 15 minutes of fluorescence irradiation are alternative, use VSV, the Sindbis indicator virus is measured the influence of fluorescence irradiation time difference to deactivation plasma viral effect, and the result is as shown in table 1 below.
Table 1 methylene blue+fluorescence irradiation different time contains VSV, the variation of Sindbis virus blood plasma log TCID50
Light application time (minute) virus titer 100ml diluted plasma 0 (containing 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
According to embodiment 1 described method, respectively following table 2 listed blood plasma are carried out inactivation of viruses and handle, the result is as shown in table 2 below.
Table 2 methylene blue/fluorescence irradiation coal fire plasma viral embodiment preface blood plasma methylene residue plasma protein response rate thrombin response rate volume indigo plant as a result goes white thin
Remove rate born of the same parents (mL) (%) (* 10
4) IgG IgM IgA Fg II VII VIII IX PLG AT-III1 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 81.25 75.32 90.123 109 ≥85.0 ≤1.0 100. 81.21 100. 87.82 78.65 100. 100. 71.21 98.21 98.654 132 ≥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 91.30 67.02 1006 116 ≥85.0 ≤1.0 98.08 88.68 94.79 73.99 78.65 87.38 98.08 73.13 85.53 90.287 106 ≥85.0 ≤1.0 100. 100. 100. 100. 100. 91.96 100. 90.74 73.17 95.458 103 ≥90.0 ≤1.0 88.69 96.84 84.27 82.74 86.42 91.67 88.69 89.83 91.84 100.9 121 ≥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
The present invention adopts photochemistry, and in conjunction with physical method, preparation is through the fresh frozen plasma of inactivation of virus, leucocyte-removing, but as killed cells is interior, extracellular virus.Whole process of preparation is carried out in disposable use, airtight blood component collection, separation, leucocyte-removing, absorption photosensitizer system, does not need sterile working's environment; More than the virus titer decline 6logTCID50, the residue leukocyte count is less than 1.0 * 10
4, the clearance of photosensitizer is greater than 85%, and the thrombin response rate is greater than 80%; Easy to operate, safe and reliable, easily apply.Describedly under the stored frozen condition of blood plasma in routine of inactivation of viruses, can preserve 2 years, greatly facilitate use clinically.
Claims (6)
1. the ablation method of a plasma viral is characterized in that it comprises: from whole blood, separate behind the blood plasma, the amount that reaches the 0.25-1.5 micromoles per liter with the ultimate density of photosensitizer in blood plasma is added photosensitizer; With 30,000-45, the fluorescence intensity irradiation of 000Lx 20-80 minute; Filtering removing leukocytes and photosensitizer, the viral blood plasma that obtained deactivation like this.
2. ablation method according to claim 1 is characterized in that described photosensitizer is selected from methylene blue, crystal violet, toluidine blue, part and secretes gland 540 and psoralen.
3. ablation method according to claim 1 is characterized in that the ultimate density of photosensitizer in blood plasma of being added is 1.0 micromoles per liter.
4. device that is used for the deactivation plasma viral, it is characterized in that it comprises blood taking needle (1), blood taking bag (3), conduit (2), (4), (5), (7), (8), (10), (12), (13) and (16), first transfering bag (6), conduction type photosensitizer easy to break adds assembly (9), fluorescence irradiation bag (11), leucocyte-removing absorption photosensitizer transfusion filter spare (15) and blood plasma reservoir bag (17).
5. device according to claim 4 is characterized in that described conduction type photosensitizer easy to break adds assembly (9) and comprises conduction type catheter adapter easy to break (20), photosensitizer liquid-feeding tube (21), liquid storage pipe (22) and closed head easy to break (23).
6. according to claim 4 or 5 described devices, it is characterized in that this device further comprises blood platelet storage bag 19, it links to each other with described device by conduit 18.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98121328 CN1112937C (en) | 1998-10-07 | 1998-10-07 | Blood plasma virus deactivating method and apparatus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98121328 CN1112937C (en) | 1998-10-07 | 1998-10-07 | Blood plasma virus deactivating method and apparatus |
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| CN1249952A true CN1249952A (en) | 2000-04-12 |
| CN1112937C CN1112937C (en) | 2003-07-02 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 98121328 Expired - Lifetime CN1112937C (en) | 1998-10-07 | 1998-10-07 | Blood plasma virus deactivating method and apparatus |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004103443A1 (en) * | 2003-05-22 | 2004-12-02 | Beijing Jingjing Medical Equipment Co., Ltd. | Method for inactivating virus in circular blood and its applictions 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 and its application |
| CN1325061C (en) * | 2002-10-16 | 2007-07-11 | 旭化成医疗株式会社 | Plasma preparation or serum preparation and process for producing the same |
| CN100413542C (en) * | 2006-06-01 | 2008-08-27 | 上海市血液中心 | 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 | Method for inactivating virus in blood product |
| 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 |
| CN104248643A (en) * | 2014-08-12 | 2014-12-31 | 中国人民解放军总医院 | Preparation device and preparation method of universal virus inactivation blood plasma |
| CN104399135A (en) * | 2014-11-20 | 2015-03-11 | 上海市血液中心 | Processing device for plasma virus inactivation |
| US20160144095A1 (en) * | 2014-11-20 | 2016-05-26 | City Of Hope | Treatment device for plasma virus inactivation |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006116899A1 (en) * | 2005-04-29 | 2006-11-09 | Chengdu Kuachang Medical Industrial Limited | The system, solid phase medium, device and method for bio-fluid treatment |
-
1998
- 1998-10-07 CN CN 98121328 patent/CN1112937C/en not_active Expired - Lifetime
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1325061C (en) * | 2002-10-16 | 2007-07-11 | 旭化成医疗株式会社 | Plasma preparation or serum preparation and process for producing the same |
| US7592134B2 (en) | 2002-10-16 | 2009-09-22 | Asahi Kasei Medical Co., Ltd. | Viral reduction method for plasma using a leukocyte-reduction filter and two virus-reduction filters of decreasing pore diameters |
| WO2004103443A1 (en) * | 2003-05-22 | 2004-12-02 | Beijing Jingjing Medical Equipment Co., Ltd. | Method for inactivating virus in circular blood and its applictions in treating viral diseases |
| 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 and its application |
| CN100413542C (en) * | 2006-06-01 | 2008-08-27 | 上海市血液中心 | 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 | Method for inactivating virus in blood product |
| 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 |
| CN104248643A (en) * | 2014-08-12 | 2014-12-31 | 中国人民解放军总医院 | Preparation device and preparation method of universal virus inactivation blood plasma |
| CN104399135A (en) * | 2014-11-20 | 2015-03-11 | 上海市血液中心 | Processing device for plasma virus inactivation |
| US20160144095A1 (en) * | 2014-11-20 | 2016-05-26 | City Of Hope | Treatment device for plasma virus inactivation |
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| CN1112937C (en) | 2003-07-02 |
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Owner name: SHANGHAI BLOOD TRANSFUSION TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: SHANGHAI CITY BLOOD CENTER Effective date: 20030905 |
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