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WO2001077045A1 - Catalyse de reaction amelioree par apport d'energie et utilisations de ladite catalyse - Google Patents

Catalyse de reaction amelioree par apport d'energie et utilisations de ladite catalyse Download PDF

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WO2001077045A1
WO2001077045A1 PCT/US2001/011703 US0111703W WO0177045A1 WO 2001077045 A1 WO2001077045 A1 WO 2001077045A1 US 0111703 W US0111703 W US 0111703W WO 0177045 A1 WO0177045 A1 WO 0177045A1
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energy
reaction
electromagnetic
molecules
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Stephen T. Flock
Kevin S. Marchitto
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    • H—ELECTRICITY
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    • B01L2300/1861—Means for temperature control using radiation

Definitions

  • the present invention relates generally to the fields of biochemical reaction and biomedical physics. More specifically, the present invention relates to methods/devices used for enhancing catalytic chemical reaction by adding energy to th e reactants therefore to increase the frequency at which reactants reach transition states of reaction.
  • Activation Energy and Reaction Rates In any given chemical reaction, the equilibrium of the reaction is given by th e difference in ⁇ G° for the reaction. The equilibrium concentrations of substrate (A) and product (C) are determined by their difference in free-energy content,
  • a catalyst lowers th e activation energy of the rate determining steps, thereby speeding up the reaction.
  • the role of the catalyst is to permit the formation of a transition state of lower energy (higher stability relative to reactants) than that for non-catalyzed reactions.
  • the catalyst itself does not participate in the reaction stoichiometry (not consumed) and cannot affect the equilibrium position of th e reaction. Pauling expressed that stabilization of the transition state of a reaction by an enzyme suggests that the enzyme has a higher affinity for the transition state than it does for th e substrate or products. ⁇ G' therefore, is reduced during catalysis.
  • the polymerase chain reaction is a technique used for in vitro and in situ amplification of specific DNA sequences.
  • the process goes in receptive cycles: denaturing, wherein the DNA of interest is denatured for about 4 minutes a t 94°C; annealing, wherein the appropriate part of the DNA strands are annealed to the primers (i.e., the antisense DNA fragment o f interest) at 50°C for about 2 minutes; and extension, wherein a heat stable enzyme called Taq-DNA-polymerase (Taq) polymerizes the individual DNA bases (deoxyribonucleotides) for 3 minutes at 72°C.
  • Taq heat stable enzyme
  • Such cycle is repeated for N times (-20- 30 times) with more primers and nucleotides added.
  • heating parameters may be modified slightly (for example, denaturing for lmin. at 94°C; annealing for 2 min . at 50°C; and extending for 3.5 min at 72°C).
  • th e heating parameters for the last cycle are also typically different, for example, denaturing for lmin. at 94°C; annealing for 2 min. a t 50°C; and extending for 10 min at 72°C.
  • the DNA o f interest is amplified by 2 N .
  • Biochemical Reaction Catalysis In order to undergo a chemical reaction, a reactant or reactants must first gain energy (activation energy) to form an activated complex before they c an proceed forward to a state (products) of different energy o r enthalpy. Generally, enzymes are used to help the reaction take place (i.e. catalyze the reaction). Heat can also be used to speed up the reaction rate. Additionally, vibrational excitation can b e used to promote endoergic (energy consuming) reactions [1].
  • the prior art is deficient in the lack of effective means of enhancing reaction catalysis by adding energy to the reactants .
  • the present invention fulfills this long-standing need and desire in the art.
  • the present invention provides methods of enhancing reaction catalysis by adding energy to reactants.
  • th e present invention provides a method of using electromagnetic energy or propagating pressure waves to enhance reaction, wherein vibrations, rotations, and/or particular configurations o r orientations of the reactants are induced. When done properly, these methods do not cause irreversible damage to the reactants and are especially suitable for use in biochemical reactions b o th in vitro and in vivo.
  • the electromagnetic energy is generated by a source which provides radiant energy with wavelength from about 200 nm to about 20,000 nm.
  • Representative examples o f electromagnetic energy include radiofrequency wave and microwave, and representative example of mechanical energy is a pressure wave.
  • the chemical reaction is a catalyzed reaction, it can be either a liquid-phase reaction or a solid-phase reaction.
  • a device for accelerating a chemical reaction comprising a reaction chamber; and a means for applying energy to the reaction chamber.
  • a method of enhancing a polymerase chain reaction by applying energy to the reaction is provided.
  • Such method c an also be used for enhancing an enzyme linked immunoassay reaction.
  • a device for enhancing a polymerase chain reaction comprising a reaction chamber; and a means for applying energy to the reaction chamber.
  • Such device can also b e used for enhancing an enzyme linked immunoassay reaction.
  • a method of increasing the rate a t which a group of molecules reaches a different molecular configuration from initial configuration comprising the step o f applying energy to the molecules.
  • the energy is electromagnetic energy or mechanical energy.
  • Representative examples of electromagnetic energy include radiofrequency wave and microwave, and representative example of mechanical energy is a pressure wave.
  • the different molecular configuration is a transition state in a chemical reaction, preferably, a catalyzed chemical reaction.
  • a device for increasing the rate at which a group of molecules reaches a different molecular configuration from initial configuration comprising a chamber for holding the molecules; and a means for applying energy to the chamber.
  • Figure 1 shows reaction coordinates as a function o f the free energy.
  • the free energy of activation, ⁇ G' is the energy that must be overcome for the reaction to proceed.
  • Figure 2 shows reaction coordinates as a function of the free energy in the presence of a catalyst.
  • the free energy o f activation, ⁇ G', is much smaller compared to that without the catalyst (see Figure 1).
  • Figure 3 shows reaction coordinates as a function of the free energy in the presence of an enzyme.
  • the free energy o f activation, ⁇ G' is broken into smaller increments by the step wise action of the enzyme catalyst which first forms an enzyme- substrate complex (i.e., ES), reaches the intermediate transition state (1), and the intermediate transition state (2), and finally proceeds to the subsequent product.
  • ES enzyme- substrate complex
  • Figure 4A shows a kind of device used for enhancing a PCR reaction.
  • Such device comprises a radiant energy source for heating the reaction; a microvessel containing the reactants ; and a cooling chamber.
  • a catalyst can be included.
  • the vessel alternates between heating and cooling cycles allowing standard PCR reaction to proceed.
  • an optional reaction chamber without the energy absorbing target/transducer, may be used to directly delivery energy to th e reactants for either local heating or for increasing the energy state of the reactants thereby increasing the frequency at which the reactants reach transition state.
  • the device 1 shown in Figure 4A comprises pulsed laser source 2 to generate a laser beam, expanding/focusing optics 3 , radiant energy absorbing target/transducer 4 to transduce propagating pres sure waves, a reaction chamber 5 containing reaction mixture and stir bars 6 , and heater/cooler 7.
  • Figures 4B-4E show that different shaped radiant energy absorbing targets within the walls of th e reaction chamber 5 , or inside the reaction solution, produc e different pressure waves.
  • Figure 5 is an energy diagram demonstrating alternating cycles of radiant energy for multi-step reactions su ch as the optically enhanced PCR.
  • Energy reaching level (2) is sufficient to separate the strands of DNA.
  • the energy is then reduced to a lower level (1) that favors a second reaction in th e sequence by catalysis.
  • Level (1) is not great enough to denature the molecular products. This cycle is repeated to produce long strands of DNA.
  • the present invention describes a device composed o f a reaction vessel where radiant energy may be supplied in such a way that reactants within the vessel are exposed to a free energy increase in the system which causes increased vibration o r rotation in molecular groupings, or causes electrons to shift t o excited states making the molecules more reactive.
  • a catalyst present in or attached to the reaction vessel in such a way th at the reactants are in contact with the catalyst, will then be in position to stabilize transition states of the reactants, thereby improving the chances that the reaction will proceed forward.
  • the reaction vessel may contain at least one reactant and one enzyme that reacts in a step-wise manner to generate products.
  • Radiant energy of a certain wavelength and energy is delivered to the reaction vessel in such a way that the reactants are activated so the first step catalytic reaction proceeds at a faster pace.
  • a second wavelength, and/or energy is then applied to the reaction such that a second step, which requires a different energy of activation, can then take place.
  • steps may b e repeated for cyclic reactions.
  • a sample o f chromosomal or other DNA is present in the reaction chamber with an oligonucleotide primer and the catalyst.
  • the first step laser radiation is added to the chamber to increase the energy o f the reaction such that the strands of the DNA are denatured and separated either locally or for the greater length of the molecule.
  • the laser energy is removed, either completely or in between pulses, the strands of DNA will then re-anneal.
  • the primer will compete for binding sites to the strand of denatured DNA. If the primer is in excess relative to the concentration of the sister DNA strand, more primer will bind as opposed to strands re-annealing.
  • radiant energy is provided t o an enzyme catalyzed reaction mixture such that energy, in th e form of heat, is delivered locally in the immediate environment o f the molecule, thereby resulting in thermal acceleration of th e rate of reaction.
  • This local heating effect is only transient, on th e order of less than microseconds, in the case of a pulsed laser. However, it is of sufficient length to result in an increase in th e frequency at which these molecules reach their transition state. In the presence of a catalyst, this transition state is stabilized and the reaction proceeds. Specifically, the use of short, high-energy pulses of radiant energy results in little heating of th e surrounding medium.
  • the electromagnetic energy is generated by a source which provides radiant energy with wavelength from about 200 nm to about 20,000 nm.
  • Representative examples o f electromagnetic energy include radiofrequency wave and microwave, and representative example of mechanical energy is a pressure wave.
  • the chemical reaction is a catalyzed reaction, it can be either a liquid-phase reaction or a solid-phase reaction.
  • a device for accelerating a chemical reaction comprising a reaction chamber; and a means for applying energy to the reaction chamber.
  • a method of enhancing a polymerase chain reaction by applying energy to the reaction is provided.
  • Such method c an also be used for enhancing an enzyme linked immunoassay reaction.
  • a device for enhancing a polymerase chain reaction comprising a reaction chamber; and a means for applying energy to the reaction chamber.
  • Such device can also b e used for enhancing an enzyme linked immunoassay reaction.
  • the energy is electromagnetic energy or mechanical energy.
  • electromagnetic energy include radiofrequency wave and microwave
  • mechanical energy is a pressure wave.
  • the different molecular configuration is a transition state in a chemical reaction, preferably, a catalyzed chemical reaction.
  • a device for increasing the rate at which a group of molecules reaches a different molecular configuration from initial configuration comprising a chamber for holding th e molecules; and a means for applying energy to the chamber.
  • Enzymatic catalysts are believed to lower ⁇ G' by replacing a single step of large ⁇ G' with several smaller steps o f lower ⁇ G' ( Figure 3).
  • enzyme mediated catalysis is extremely efficient for reactions with large ⁇ G' because each incremental step is composed of a smaller transition energy o f activation.
  • the step wise transitions may be taken advantage of and the undesirable reactions and denaturation avoided.
  • a continuous wave or pulsed laser is used to heat the surrounding medium rapidly in such a way th at the heat involved in the reaction increases rapidly.
  • a small reaction volume can then be cooled rapidly so that unstable products or side reactions are minimized.
  • This approach is particularly useful when cyclic reactions are necessary, wherein rapid heating and cooling steps may take place in relatively small volumes.
  • Energy may also be added to the system using microwaves or radiofrequency waves in the same manner .
  • Radiofrequency waves are of particular interest because they are capable of causing the vibrational effects in the medium without heating of the applicator. In effect, radiofrequency waves may b e incorporated into a heating element that does not heat up the element itself during use.
  • the heating element could always be kept cold while it transmits energy to the reaction medium in direct contact with or near to it.
  • short and repeated pulses of radiant energy are directed at the molecular species which causes a n increased level of excitation.
  • This approach takes advantage o f the fact that molecular species absorb photonic energy, which increases their intrinsic rate of rotation or vibration and th e likelihood that inner shell electrons will be promoted to outer shells, thereby creating a less stable structure.
  • the above described methods can also be used for improving reaction rates in solid-phase reactions.
  • a method/device is hereby provided to increase th e speed and efficiency of PCR. Such method/device may also increase the yield of PCR products and reduce the amount o f reactants used by virtue of minimizing or eliminating heating o f the PCR reaction solution. Reducing or eliminating the heat involved in denaturation also reduces or eliminates the need for thermostable enzymes. Strategies described within include activating molecules or molecular groups through energy absorption from infrared radiation. Absorption of infrared radiation by biomolecules can be broken down into three regions.
  • an OH group stretching vibration is near 7140 cm” 1 (1.4 microns) and an NH stretching vibration is near 6667 cm 1 (1.5 microns).
  • 4000 to 1300 cm “ 1 (2.5-7.7 microns) is the "group frequency region"
  • 1300 to 650 cm 1 (7.7-15.4 microns) is the "fingerprint region”.
  • Absorptions depend on the functional group present (in the case of the former) and not the complete molecular structure, or depend on single-bond stretching and bending vibrations (in the case of the latter).
  • Denaturation of DNA can be achieved using radiative energy [2].
  • Radiant energy with a wavenumber of about 1550- 1800 cm 1 induces in-plane double-bond vibrations of bases, with a wavenumber of about 1275- 1550 cm " 1 induces base deformation motions coupled through glycosidic linkages t o sugar vibrations, with a wavenumber of about 1050- 1275 cm 1 induces phosphate and sugar vibrations, and with a wavenumber of about 750-1050 cm 1 induces vibrations of the sugar- phosphate backbone and bases. All of these infrared spectral domains are in the infrared region of the electromagnetic spectrum (wavelengths of about 5.5-12.5 microns).
  • the 1225 cm '1 and 1084 cm - 1 (asymmetric and symmetric stretching vibrations o f nucleic acids) bands could be targeted by lasers emitting long- wavelength radiant energy; in the case of the former, a CO 2 laser would suffice.
  • DNA has hydrogen bonds between the two strands .
  • hydrogen bonds For every adenine and thymine base, there are two hydrogen bonds, viz.: O- - -H and N - H.
  • the ionic OH and NH bonds have energies on the order of 111 and 9 3 kcal/mol respectively, while the O- - -H and N - H hydrogen bonds have energy on the order of about 1 kcal/mol, which is why they are easily broken.
  • One method of breaking hydrogen bonds is by continued exposure to temperatures on the order o f 75°C.
  • hydrogen bonds can be broken by radiant energy photons with a wavelength of about 32 microns or less.
  • the ionic OH and NH bonds require photons with wavelengths of about 307 nm or less. Consequently, one c an break hydrogen bonds in DNA without causing breaks in the DNA strands themselves (i.e.
  • the OH stretching vibration near 1.4 microns and the NH stretching vibration near 1.5 microns could be excited with radiant energy produced by, for example, an Nd:YLF or Nd: glass laser.
  • radiant energy produced by, for example, an Nd:YLF or Nd: glass laser.
  • irradiating the DNA with 1.4-1.5 micron radiant energy prior to the hydrogen-bond-breaking step done with radiant energy at a longer wavelength would increase the reaction rate.
  • by irradiating the denatured DNA with 1 .4- 1 .5 micron radiant energy after the denaturation step would increase the reaction rates of the annealing and extension step.
  • different wavelengths may be used to differentially o r combinatorially enhance the conversion of species from one t o another.
  • Compressional or tensile propagating pressure waves can be produced by irradiating biological media with pressure waves produced by acoustic transducers (such a s piezoelectric crystals or ultrasonic transducers) or radiant energy produced by a pulsed laser, or by irradiating absorbing materials placed in intimate acoustic contact with tissue of interest.
  • Shock waves propagating pressure waves moving in a medium a t velocities greater than the speed of sound in the medium
  • a propagating pressure wave can transform into a shock wave within a short distance while propagating in water, for example.
  • a pressure wave with an amplitude of 300 bars can provide about 0.72 kcal/mol of energy. This is sufficient to break hydrogen bonds, but not enough to break covalent or ionic bonds.
  • the ability to break bonds and induce vibrations results from not only the amplitude of the wave, b u t also the temporal characteristics. Further, the addition of a n appropriate catalyst to any of these enhanced reactions would favor stabilization of transition molecules and therefore further increase the forward reaction rate.
  • FIG. 4A When propagating pressure waves are produced b y pulsed laser radiant energy impinging on an absorbing target ( Figure 4A), different shaped targets may be beneficial ( Figures 4B-4E). For example, different shaped targets within the walls o f the reaction chamber, or inside the reaction solution, c an produce pressure waves that can be beneficial. Multiple lasers can also be used to produce multiple propagating pressure waves impinging on the reaction mixture from different directions. It is beneficial to continually mix the reaction mixture and place th e reaction chamber on a controllable heating/cooling element t o maintain the temperature of the reaction solution in a region suitable for the process to take place.
  • reaction mixture it can be beneficial to de-gas the reaction mixture, or introduce hyperbaric inert gases (e.g. N 2 ) in the reaction mixture, so as to maintain a sealed reaction vessel in order to modulate the extent to which cavitation bubbles are produced.
  • hyperbaric inert gases e.g. N 2
  • these bubbles propagate and carry significant amounts of energy to damage biological media, such as DNA.
  • they may b e used to beneficially add energy to the reaction mixture.
  • pulsed or continuous wave radiant energy c an be used to directly excite molecules to which light-absorbing complexes are attached.
  • energy can be added to the reactants by transference from these dye-conjugates molecules or simply by the dye itself upon the absorption of a pulse of radiant energy from, for example, an alexandrite laser tuned to emit energy with a wavelength of 795 nm.
  • a pulse of radiant energy from, for example, an alexandrite laser tuned to emit energy with a wavelength of 795 nm.
  • Catalytic reactions often rely on specific interactions among various reactants or between the catalyst and reactants .
  • the shape and/or size of molecules can sometimes determine where the molecule has an affinity to localize and bind.
  • a particular protein can interact with a particular target molecule when the protein is folded into it's "native" state; when unfolded, or denatured, it typically cannot interact with th e target molecule in spite of the fact that the protein still has the same peptide sequence.
  • the particular desirable (or deleterious) protein- protein, protein-DNA or protein-molecule interaction depends o n the correct binding of the molecules.
  • the efficiency of binding o r reacting can be increased by increasing the rate at which th e molecules "bump" into each other. This is typically a function o f concentration and kinetic energy in the system.
  • binding or reacting can be altered by a change in conformation o f one or both of the molecules.
  • a conformational change may increase the exposure of the binding sites on each molecule o r may alter the shape of each molecule such that they c an approach each other more closely. For example, it is generally accepted that blocking the flow of sodium through the sodium channel in nerve cell membranes makes the nerve inexcitable by local action currents.
  • Changes in the shape and caliber of this channel determines what anesthetics are efficacious.
  • the shape and caliber of the channel can be changed by, for example, action potentials, toxins, and local anesthetics that bind to gating receptor sites. Some local anesthetics bind to a channel protein binding site thus rendering the sodium channel blocked.
  • Transport of molecules across membranes or o ther tissue barriers may be enhanced through application of th e concepts described herein.
  • irradiation of a membrane or the skin will result in transient changes in the molecular configuration of its constituents. Molecules which come into contact with the irradiated molecules will have a n accelerated rate of binding or reacting as compared to those in non-irradiated tissue.
  • This concept may be applied to drug permeation through membranes or tissue barriers.
  • one of the rate limiting steps in permeation of compounds through membranes or tissue barriers is the rate of initial binding to local receptors.
  • anesthetics such as lidocaine
  • permeation is in part related to the rate of local binding to receptors.
  • a reaction involving test molecules is enhanced by transiently and repetitively raising th e temperature of the surrounding environment (i.e. the ambient molecules) .
  • PCR reaction A PCR mixture including buffers, primers, Taq polymerase and deoxyribonucleotides was prepared as per manufacturer' s instructions and placed in a reaction vial. Equal amount of a sample of DNA to be amplified was added t o the mixture. The reaction mixture was exposed to the radiant energy produced by a Ho:YAG laser. The laser radiant energy output was configured to produce a spot size at the reaction vial of 5 mm, with a 400 microsecond pulse, 10 Hz pulse repetition rate, and 200 mJ/pulse pulse energy. The radiant energy, which was absorbed by the reaction mixtures, was arranged to impinge on the lateral side of the reaction vial in a position so that the reaction mixture was irradiated.
  • ELISA An enzyme linked immunoassay (ELISA) reaction mixture was prepared in 96-well plates previously seeded with equal numbers of rat cancer cells. The reaction mixture was MTT, which produces a colorimetric change in th e presence of viable cells. After adding the assay reaction mixture to the wells in the plate, single wells were irradiated with the radiant energy produced by a Ho:YAG laser.
  • the laser radiant energy output which was absorbed by the reaction mixtures, was configured to produce a spot size at the surface of the reaction mixture of 5 mm, with a 400 microsecond pulse, 10 Hz pulse repetition rate, and 200 mJ/pulse pulse energy.
  • the irradiation continued on for a period of 10 minutes, after which a solvent was added and the mixture was spectroscopically assayed for color.
  • the result shows that a greater degree of color change (in other words, a greater optical density OD) occurred in th e irradiated mixture as compared to controls. Since the wells were seeded with equal numbers of cells, the greater OD is indicative of an enhanced reaction catalysis and not of different numbers o f viable cells.
  • an ELISA reaction mixture was prepared in 96-well plates previously seeded with equal numbers of ra t cancer cells.
  • the reaction mixture was MTT, which produces a colorimetric change in the presence of viable cells.
  • entire 96-well plate was exposed to microwave energy by placing the plate within a 500 Watt microwave oven.
  • the microwave irradation conditions were as follows: the oven was set to the defrost mo de (lowest power possible) and the oven was sequenced on and off repetitively (approximately 2 seconds on and 10 seconds off) fo r a period of 10 minutes. After the irradiation, a solvent was added and the mixture was spectroscopically assayed for color.
  • reaction mixture a reaction involving te s t molecules is enhanced by transiently and repetitively raising th e temperature of an absorbing species added to the surrounding environment (reaction mixture).
  • a PCR mixture was prepared as per manufacturer' s instructions and placed in a reaction vial. Equal amount of a sample of DNA to be amplified was added to the mixture. The enzyme present in the reaction mixture was Taq polymerase. The reaction mixture was exposed to the radiant energy produced by a ruby laser. The laser radiant energy output was configured t o produce a spot size at the reaction vial of 5 mm, with a 400 microsecond pulse, 10 Hz pulse repetition rate, and 200 mJ/pulse pulse energy. Indocyanine green (ICG) was solubilized in ultrapure water and added to the reaction mixture to a final concentration of 1%.
  • ICG Indocyanine green
  • the radiant energy which was absorbed primarily by the ICG, was arranged to impinge on the lateral side of the reaction vial in a position so that the reaction mixture was irradiated.
  • the irradiation continued on for a period of 2 0 minutes, after which the mixture was assayed for DNA amplification.
  • the degree of amplification was compared to non- irradiated (control) samples. The result shows that greater amplification occurred in the irradiated mixture as compared t o controls .
  • a PCR mixture including buffers, primers, Taq polymerase and deoxyribonucleotides was prepared as p e r manufacturer' s instructions and placed in 0.1 mm pathlength optical cuvettes. Equal amount of a sample of DNA to b e amplified was added to the mixture. The reaction mixture was exposed to the radiant energy produced by an Nd:YLF laser. The laser radiant energy output was configured to produce a spot size at the lateral side of the cuvette of 5 mm, with a 400 microsecond pulse, 10 Hz pulse repetition rate, and 20 mJ/pulse pulse energy.
  • a reaction involving test molecules is enhanced by transiently and repetitively exposing the test molecules and reaction mixture to propagating pres sure waves.
  • a PCR mixture including buffers, primers, Taq, o r other DNA polymerase, and deoxyribonucleotides was prepared as per manufacturer' s instructions and placed in a reaction cuvette. Equal amount of a sample of DNA to be amplified was added to the mixture.
  • a Q-switched or mode-locked laser e.g. , ruby or Nd:YAG
  • a 20 ns pulse duration, 1 mm spot size, 1 0 Hz pulse repetition rate and 20 mJ pulse energy was directed on a light absorbing material (e.g. thermoelectrically cooled black anodized aluminum) in intimate acoustic contact with th e reaction cuvette.

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  • Electromagnetism (AREA)
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Abstract

L'invention concerne un procédé/dispositif (1) permettant d'améliorer une réaction chimique notamment PCR et ELISA par utilisation d'énergie électromagnétique ou mécanique. On peut également utiliser ce procédé/dispositif (1) pour augmenter la vitesse à laquelle un groupe de molécules induit une configuration moléculaire différente de la configuration initiale, ce qui permet d'accélérer la liaison et la réaction desdites molécules.
PCT/US2001/011703 2000-04-10 2001-04-10 Catalyse de reaction amelioree par apport d'energie et utilisations de ladite catalyse Ceased WO2001077045A1 (fr)

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US09/546,065 US6673214B1 (en) 1999-04-09 2000-04-10 Energy enhanced reaction catalysis and uses thereof
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002008455A3 (fr) * 2000-07-25 2002-08-15 Biotix Gmbh Procede de separation d'acides nucleiques a brin double se trouvant dans une solution, en acides nucleiques a un seul brin
WO2022020955A1 (fr) * 2020-07-31 2022-02-03 12198703 Canada Ltd. Cinétique chimique à orientation dirigée

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5689008A (en) * 1996-02-02 1997-11-18 United Technologies Corporation Catalytic reaction rate enhancement at low temperatures
WO1998006876A1 (fr) * 1996-08-16 1998-02-19 Pharmacia Biotech Inc. Dispositif et procedes destines a une conversion thermique par un transducteur induite a distance dans des reactions chimiques et biochimiques

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5689008A (en) * 1996-02-02 1997-11-18 United Technologies Corporation Catalytic reaction rate enhancement at low temperatures
WO1998006876A1 (fr) * 1996-08-16 1998-02-19 Pharmacia Biotech Inc. Dispositif et procedes destines a une conversion thermique par un transducteur induite a distance dans des reactions chimiques et biochimiques

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002008455A3 (fr) * 2000-07-25 2002-08-15 Biotix Gmbh Procede de separation d'acides nucleiques a brin double se trouvant dans une solution, en acides nucleiques a un seul brin
WO2022020955A1 (fr) * 2020-07-31 2022-02-03 12198703 Canada Ltd. Cinétique chimique à orientation dirigée
JP2023537701A (ja) * 2020-07-31 2023-09-05 １２１９８７０３ カナダ エルティーディー． 指向性化学反応速度論
US12151224B2 (en) 2020-07-31 2024-11-26 12198703 Canada Ltd. Directed orientation chemical kinetics

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