WO1996000091A1 - A method of virus inactivation in samples by uv irradiation - Google Patents
A method of virus inactivation in samples by uv irradiation Download PDFInfo
- Publication number
- WO1996000091A1 WO1996000091A1 PCT/EP1995/002444 EP9502444W WO9600091A1 WO 1996000091 A1 WO1996000091 A1 WO 1996000091A1 EP 9502444 W EP9502444 W EP 9502444W WO 9600091 A1 WO9600091 A1 WO 9600091A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- laser
- virus
- wavelength
- active components
- inactivation
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 30
- 241000700605 Viruses Species 0.000 title claims abstract description 26
- 230000002779 inactivation Effects 0.000 title claims abstract description 17
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 title 1
- 230000000694 effects Effects 0.000 claims abstract description 5
- 230000005670 electromagnetic radiation Effects 0.000 claims abstract description 4
- 230000001678 irradiating effect Effects 0.000 claims abstract description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 10
- 230000005855 radiation Effects 0.000 claims description 7
- 150000001875 compounds Chemical class 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 210000004369 blood Anatomy 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 108020004707 nucleic acids Proteins 0.000 claims description 3
- 102000039446 nucleic acids Human genes 0.000 claims description 3
- 150000007523 nucleic acids Chemical class 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 102000008072 Lymphokines Human genes 0.000 claims description 2
- 108010074338 Lymphokines Proteins 0.000 claims description 2
- 241000124008 Mammalia Species 0.000 claims description 2
- 229920001222 biopolymer Polymers 0.000 claims description 2
- 230000023555 blood coagulation Effects 0.000 claims description 2
- 239000003102 growth factor Substances 0.000 claims description 2
- 229940088597 hormone Drugs 0.000 claims description 2
- 239000005556 hormone Substances 0.000 claims description 2
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 230000003612 virological effect Effects 0.000 claims description 2
- 230000004952 protein activity Effects 0.000 claims 1
- 230000000415 inactivating effect Effects 0.000 abstract 1
- 239000013078 crystal Substances 0.000 description 6
- 230000003287 optical effect Effects 0.000 description 5
- 241000991587 Enterovirus C Species 0.000 description 2
- 108010054218 Factor VIII Proteins 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 239000010836 blood and blood product Substances 0.000 description 2
- 229940125691 blood product Drugs 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010076282 Factor IX Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 108010094028 Prothrombin Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000012414 sterilization procedure Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/22—Testing for sterility conditions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
- A61L2/0011—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using physical methods
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/08—Radiation
- A61L2/10—Ultraviolet radiation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
- C12N7/04—Inactivation or attenuation; Producing viral sub-units
Definitions
- the invention is concerned with a method of virus inacti ⁇ vation in samples having biologically active components by irradiating the sample with electro-magnetic radiation, and a device for carrying out the method of the invention.
- the object of the invention is to provide a method for virus inactivation in samples having biologically active components derived from potentially virus infected sources.
- the method should efficiently inactivate the virus but leave intact as much as possible the activity of the protein.
- the method should be applicable also in industrial scale and for various virus inactivation problems with different biological sources.
- the objects addressed above are solved by a method of virus inactivation in samples having biologically active components derived from potentially virus infected sources by differen ⁇ tially irradiating the sample or sources with electro-magne ⁇ tic radiation generated by a tuneable laser-device for inactivation the virus but not the activity of the biologi ⁇ cally active components.
- the wavelength of the electro-magnetic radiation generated by the laser-device should be in the range of from 300 to 370 nm generated by a laser-device which has tunable wavelength.
- the virus was inactivated at an inactivation level larger than 4log 10 with the remaining protein of the sample of higher than 80 %.
- the method of the invention provides a satisfying virus inactivation even at pump rates larger than 1 ml/s.
- factor VIII containing solutions are irridiated with radiation having a wavelength from 305 to 315 nm and IgG from 350 to 360 nm.
- the energy irridiated into the sample is to be larger than 3 J/ml, preferably between 10 to 20 J/ml.
- the upper limit depends on the sensitivity of the active component of interest in the solution to be virus inactivated.
- the potentially virus infected source having biologically active components is a body liquid from mammals including man which body liquid is such as whole blood, plasma, liquor cerebrospinalis, sperms, lymphe and the like.
- the compounds having biologically components are preferably high molecular compounds e.g. biopolymers such as proteins, nucleic acid, or low molecular compounds such as hormones, growth factors, lymphokines and the like.
- the proteins are preferably proteins having a function such as enzymes, antibodies, proteinfactors for regulating physio ⁇ logical cycles such as the bloodclotting cascade, e.g. Factor II, VIII, IX and the like,
- a laser is used the wavelength of which is tunable and which radia ⁇ tion can be pulsed.
- the laser-devices used according to the method of the in ⁇ vention comprises a tunable solid state laser system deli ⁇ vering energetic UV pulses into a flow cell, which is a preferred device for leading the sample to be laser-irridia- ted through the laser-beam.
- a protein solution which can be derived from a separation column eluate which is passing in * the flow cell can be pumped at a rate of 1 to 2 ml/s.
- the preferred laser device is a solid state laser, which produces currently radiation at discrete wavelength or over very limited ranges of wavelengths.
- the generation of high power and wavelength tunable radiation in the UV region is obtained by using two closely related non linear optical techniques: Optical Parametric Oscillation (OPO) and Second Harmonic Generation (SHG) .
- OPO Optical Parametric Oscillation
- SHG Second Harmonic Generation
- OPO works as schematically outlined below. Basically, the output of a high power pulse laser (the pump) is focused into a nonlinear crystal, and provided that the pump intensity is sufficient high, the optical parametric process is in ⁇ itiated, creating "signal" and "idler” beams whose optical frequencies sum up to that of the pump.
- the crystal In order to reduce the threshold for the onset of the parametric oscillation, the crystal is usually enclosed in an optical cavity which provides feedback. Tunability arises from phase matching constraints upon the three interacting waves in the crystal (pump, signal and idler) which determine the signal and idler wavelengths for a given propagation direction and temperatu ⁇ re. By changing the crystal angle or its temperature the signal and idler wavelengths shift to maintain the phase matching conditions.
- a Q switched Nd:YAG laser is the pump source.
- the pump wavelength might be either the fundamental or a harmonic of 1.064 ⁇ m.
- a variety of nonlinear materials (BBO, KTP etc.) have been demonstrated to generate radiation over the entire range from 0.3 ⁇ m to 18 ⁇ m.
- a frequency doubling crystal (Second Harmonic generation) the lower end of this range can be extended to roughly 200 nm.
- Conversion efficiencies of around 10 % to the UV can be achieved in a conversion scheme where the third harmonic of a YAG laser is downconverted to the 480 to 700 nm band and then frequency doubled to the required spectral range.
- Output pulse energies of around 50 J at 20 pps can be obtained in a large UV tuning range, with well proven and commercially available lasers and nonlinear crystals.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Virology (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
A method of virus inactivation in samples having biologically active components derived from potentially virus infected sources by differentially irradiating the sample or sources with electro-magnetic radiation generated by a tuneable laser-device for inactivating the virus but not the activity of the biologically active components.
Description
A method of virus inactivation in samples by ϋV irradiation
The invention is concerned with a method of virus inacti¬ vation in samples having biologically active components by irradiating the sample with electro-magnetic radiation, and a device for carrying out the method of the invention.
Matthews et al., Blood cells, 18, 1, (1992), pages 75 to 89 report about direct and indirect irradiation methods as a potential method for blood products sterilization. Indirect methods typically use chromophore additives, which bind to the virus, and which being strongly absorbing serve as an energy relay to the virus leading to its thermal degradation and destruction. The disadvantage associated with such a method is the use of chemical additives which imply an additional purification step for their removal following the sterilization procedure. The direct irradiation method uses light sources which correspond in their wavelengths to the absorption band of the viral species in the UV part of the spectrum. Because of overlapping of the optimum inactivation conditions and the inactivation of proteins satisfactory results could not yet achieved.
Prodouz et al. report in Blood Vol. 70, pp 589 - 592 (1987) that also direct methods were employed using a radiation at 308 nm, which were emitted from a XeCl eximer laser. The
reported inactivation of viruses in blood products containing poliovirus, platelets, and plasma. The damage on platelets* and plasma proteins was reported to be not very severe. A poliovirus inactivation of 4log10 to 61og10 was obtained. The used laser was pulsed having an intensity of lower than 170 k /cm2. The pump rate was reported to be in the order of 1 ml/s. A drawback of this method is that the difference in the absorption spectrum of protein and nucleic acid in the near TJV range is not so profound as to give a good differen¬ tiation.
The object of the invention is to provide a method for virus inactivation in samples having biologically active components derived from potentially virus infected sources. The method should efficiently inactivate the virus but leave intact as much as possible the activity of the protein. The method should be applicable also in industrial scale and for various virus inactivation problems with different biological sources.
The objects addressed above are solved by a method of virus inactivation in samples having biologically active components derived from potentially virus infected sources by differen¬ tially irradiating the sample or sources with electro-magne¬ tic radiation generated by a tuneable laser-device for inactivation the virus but not the activity of the biologi¬ cally active components.
Preferably, the wavelength of the electro-magnetic radiation generated by the laser-device should be in the range of from 300 to 370 nm generated by a laser-device which has tunable wavelength. Surprisingly, the use of shorter wavelength than 370 nm, preferably 305 nm to 355 nm, yielded a differentia¬ tion between protein content of the sample and virus. The virus was inactivated at an inactivation level larger than 4log10 with the remaining protein of the sample of higher than 80 %. Preferably, the method of the invention provides
a satisfying virus inactivation even at pump rates larger than 1 ml/s.
The actual wavelength for optimum virus inactivation can be investigated by tuning the laser's wavelength. For example, factor VIII containing solutions are irridiated with radiation having a wavelength from 305 to 315 nm and IgG from 350 to 360 nm.
The energy irridiated into the sample is to be larger than 3 J/ml, preferably between 10 to 20 J/ml. The upper limit depends on the sensitivity of the active component of interest in the solution to be virus inactivated.
Preferably, the potentially virus infected source having biologically active components is a body liquid from mammals including man which body liquid is such as whole blood, plasma, liquor cerebrospinalis, sperms, lymphe and the like.
The compounds having biologically components are preferably high molecular compounds e.g. biopolymers such as proteins, nucleic acid, or low molecular compounds such as hormones, growth factors, lymphokines and the like.
The proteins are preferably proteins having a function such as enzymes, antibodies, proteinfactors for regulating physio¬ logical cycles such as the bloodclotting cascade, e.g. Factor II, VIII, IX and the like,
In a preferred embodiment of the present invention a laser is used the wavelength of which is tunable and which radia¬ tion can be pulsed.
The laser-devices used according to the method of the in¬ vention comprises a tunable solid state laser system deli¬ vering energetic UV pulses into a flow cell, which is a preferred device for leading the sample to be laser-irridia-
ted through the laser-beam. A protein solution which can be derived from a separation column eluate which is passing in* the flow cell can be pumped at a rate of 1 to 2 ml/s. The preferred laser device is a solid state laser, which produces currently radiation at discrete wavelength or over very limited ranges of wavelengths. The generation of high power and wavelength tunable radiation in the UV region is obtained by using two closely related non linear optical techniques: Optical Parametric Oscillation (OPO) and Second Harmonic Generation (SHG) .
OPO works as schematically outlined below. Basically, the output of a high power pulse laser (the pump) is focused into a nonlinear crystal, and provided that the pump intensity is sufficient high, the optical parametric process is in¬ itiated, creating "signal" and "idler" beams whose optical frequencies sum up to that of the pump. In order to reduce the threshold for the onset of the parametric oscillation, the crystal is usually enclosed in an optical cavity which provides feedback. Tunability arises from phase matching constraints upon the three interacting waves in the crystal (pump, signal and idler) which determine the signal and idler wavelengths for a given propagation direction and temperatu¬ re. By changing the crystal angle or its temperature the signal and idler wavelengths shift to maintain the phase matching conditions.
Preferably, a Q switched Nd:YAG laser is the pump source. Depending on the OPO output wavelength, the pump wavelength might be either the fundamental or a harmonic of 1.064 μm. A variety of nonlinear materials (BBO, KTP etc.) have been demonstrated to generate radiation over the entire range from 0.3 μm to 18 μm. By using a frequency doubling crystal (Second Harmonic generation) the lower end of this range can be extended to roughly 200 nm. Conversion efficiencies of around 10 % to the UV can be achieved in a conversion scheme where the third harmonic of a YAG laser is downconverted to
the 480 to 700 nm band and then frequency doubled to the required spectral range. Output pulse energies of around 50 J at 20 pps can be obtained in a large UV tuning range, with well proven and commercially available lasers and nonlinear crystals.
Claims
1. A method of virus inactivation in samples having biolo¬ gically active components derived from potentially virus infected sources by differentially irradiating the sample or sources with electro-magnetic radiation generated by a tuneable laser-device for inactivation the virus but not the activity of the biologically active components.
2. The method of claim 1, wherein the potentially virus infected source having biologically active components is a body liquid from mammals including man which body liquid is such as whole blood, plasma, liquor cerebro- spinalis, sperms, and lymphe.
3. The method of claim 1 and/or 2, wherein the biological¬ ly active components are high molecular compounds e.g. biopolymers such as proteins, nucleic acids, or low molecular compounds such as hormones, growth factors and lymphokines.
4. The method of claim 3, wherein the proteins are pro¬ teins having a function such as enzymes, antibodies, proteinfactors for regulating physiological cycles such as the blood-clotting cascade.
5. The method of any one of the claims 1 to 4, wherein the wavelength of the laser radiation would be in the range of from 300 nm to 370 nm if the sample contains active proteins the activity of which has to be preserved.
6. The method of claim 5, wherein the wavelength is in the wavelength range of from 305 to 355 nm.
7. The method of claim 6, wherein a pulsed laser device is used.
8. The method of anyone of the claims 1 to 7, wherein a viral inactivation level larger than 4 log10 and a remaining protein activity of higher than 80 % is achieved.
9. The method of any one of the claims 1 to 8, wherein the the pump rate for the transportation of the samples to be treated is larger than 1 ml/s.
10. A device for carrying out the method of any one of the claims 1 to 9, comprising a laser device having a laser beam source, devices for tuning the wavelength in the UV range, and a device for leading the sample to be laser-irradiated through the laser-beam.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU28874/95A AU2887495A (en) | 1994-06-25 | 1995-06-23 | A method of virus inactivation in samples by uv irradiation |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP94109862 | 1994-06-25 | ||
| DE94109862.6 | 1994-06-25 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1996000091A1 true WO1996000091A1 (en) | 1996-01-04 |
Family
ID=8216053
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/EP1995/002444 WO1996000091A1 (en) | 1994-06-25 | 1995-06-23 | A method of virus inactivation in samples by uv irradiation |
Country Status (2)
| Country | Link |
|---|---|
| AU (1) | AU2887495A (en) |
| WO (1) | WO1996000091A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000047240A1 (en) * | 1999-02-13 | 2000-08-17 | Purepulse Technologies, Inc. | Methods of inactivating pathogens using broad-spectrum pulsed light |
-
1995
- 1995-06-23 AU AU28874/95A patent/AU2887495A/en not_active Abandoned
- 1995-06-23 WO PCT/EP1995/002444 patent/WO1996000091A1/en active Application Filing
Non-Patent Citations (2)
| Title |
|---|
| J.L. MATTHEWS ET AL.: "Inactivation of viruses with photoactive compounds", BLOOD CELLS, vol. 18, NEW YORK NY USA, pages 75 - 89 * |
| K.N. PRODOUZ ET AL.: "Use of laser-UV for the inactivation of virus in blood products.", BLOOD, vol. 70, no. 2, 1 August 1987 (1987-08-01), WASHINGTON DC USA, pages 589 - 592 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000047240A1 (en) * | 1999-02-13 | 2000-08-17 | Purepulse Technologies, Inc. | Methods of inactivating pathogens using broad-spectrum pulsed light |
| AU779193B2 (en) * | 1999-02-13 | 2005-01-13 | Purepulse Technologies, Inc. | Methods of inactivating pathogens using broad-spectrum pulsed light |
| KR100630520B1 (en) * | 1999-02-13 | 2006-09-29 | 퓨어펄스 테크놀러지즈 인코포레이티드 | Methods of inactivating pathogens using broad-spectrum pulsed light |
Also Published As
| Publication number | Publication date |
|---|---|
| AU2887495A (en) | 1996-01-19 |
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