18 results about "Thermal denaturation" patented technology

The invention discloses a biosensor based on an ultraviolet light triggered bio-barcode release technology (OBCA) and construction and application of the biosensor. The biosensor comprises nucleic acid functionalized silver nanoparticles, nucleic acid functionalized magnetic beads and a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas12a detection system, the DNA bar code modified on the surface of the silver nanoparticle contains a light cleavable linker, a CRISPR / Cas12a activation sequence and a target capture sequence I, and a DNA capture probe containing a target capture sequence II is fixed on the surface of the magnetic bead through biotin-streptavidin action. During detection, target miRNA mediates two capture sequences to form a sandwich compound, after magnetic separation and thermal denaturation elution, 365nm ultraviolet radiation releases DNA bar code fragments, and the Cas12a trans-cleavage fluorescence quenching reporter probe is activated to generate a signal. According to the method, nucleic acid amplification and reverse transcription are not needed, the detection limits of miR-21, miR-155 and mi-210 respectively reach 158.94 aM, 763.56 aM and 197.82 aM, single-base mismatch and precursor miRNA can be distinguished, the serum sample detection result is highly consistent with that of RT-PCR, and the method is suitable for high-sensitivity quantitative detection of miRNA.

This invention relates to the field of biomedical materials, providing non-denatured type II collagen ultrafine powder with cartilage tissue-targeted growth and structural repair functions, its bioactivity preservation preparation method, and its application in joints. The ultrafine powder has an average particle size of 0.5-50 μm, a triple helix structure integrity rate of not less than 85%, a thermal denaturation temperature of 38-48℃, and retains chondrocyte binding domains CB10, CB11, and MMP cleavage sites. The preparation method includes raw material pretreatment, time-limited enzymatic hydrolysis, graded purification, and ultra-low temperature ball milling. The product can be used for the repair and treatment of cartilage degenerative diseases such as osteoarthritis and cartilage damage.

The invention relates to the technical field of biomedical materials, in particular to polynucleotide / macromolecular protein composite gel capable of tolerating terminal moist heat sterilization as well as a preparation method and application of the polynucleotide / macromolecular protein composite gel. The product provided by the invention is a composite gel of polynucleotide PN and a specific type of high-molecular protein, and under the conditions of a specific ratio and molecular weight, the high-thermal denaturation temperature is higher, and the high-molecular protein with a specific structure has a better protection effect on the polynucleotide PN; the stability and biological activity of polynucleotide PN molecules can be kept under the condition of high-temperature terminal sterilization, the safety of a sterilized product can achieve the effect of thoroughly inactivating viruses, and the filling and supporting properties of the product serving as a stent material are completely reserved; the compound can be directly used in drugs, medical instruments or cosmetics in the field of tissue engineering, and has high safety and lasting effect.

The invention relates to the technical field of micro-fluidic electrochemical detection, in particular to a multi-channel micro-fluidic chip for detecting various drug-resistant mycobacterium tuberculosis as well as a use method and application of the multi-channel micro-fluidic chip. The multi-channel micro-fluidic chip comprises a splitting area and a sample mixing area, the splitting area comprises a sample injection hole and a DNA splitting chamber which are communicated in sequence, the sample mixing area comprises a sample mixing chamber, and the DNA splitting chamber and the sample mixing chamber are communicated through a first micro-channel; the electrode array area comprises a plurality of reaction detection chambers and detection electrodes, the detection electrodes are in one-to-one correspondence with the reaction detection chambers and are in contact with the inner spaces of the reaction detection chambers, the electrodes are modified with signal amplification probes, Cas12a-crRNA compounds are preloaded on the surfaces of the electrodes, and the reaction detection chambers are communicated with the sample mixing chamber through a second micro-channel. Multiple channels are designed, synchronous detection of multiple drug-resistant genes is achieved, integration of clinical sample treatment, DNA acquisition, DNA thermal denaturation treatment and DNA detection is achieved, operation is easy and smooth, and clinical diagnosis and medication guidance of tuberculosis drug resistance can be well served.

The invention discloses differential temperature comparison single molecule detection based on a reversible thermal change renaturation query probe, and relates to the field of single molecule immunodiagnosis. According to the invention, the property of repeated thermal denaturation / renaturation of the query probe is utilized, and the specific binding activity of the query probe is controlled, so that a specific binding molecular signal and a non-specific binding molecular signal of the query probe are distinguished, and a novel high-specificity single-molecule immunodiagnosis method is provided.

This application discloses an ultrasonic thermal strain measurement method and system based on equivalent thermal dose correction, relating to the field of thermotherapy monitoring: acquiring the initial temperature of biological tissue; during the heating process of biological tissue, acquiring ultrasonic echo signal sequences of the target area at preset time intervals; calculating the cumulative thermal strain of the target area at each measurement moment based on the ultrasonic echo signal sequence; coupling and predicting the temperature and equivalent thermal dose at the current moment based on the equivalent thermal dose and temperature at the previous measurement moment, obtaining the predicted values ​​of the temperature and equivalent thermal dose at the current moment; establishing a temperature inversion equation including cumulative thermal strain and equivalent thermal dose, iteratively solving the equation using the predicted value as the initial value, and outputting the corrected temperature value and updated equivalent thermal dose value at the current moment. This application achieves dynamic compensation for the nonlinear changes in acoustic parameters caused by tissue thermal denaturation, thereby improving the temperature measurement accuracy during the ablation stage, expanding the temperature measurement range, and realizing real-time synchronous monitoring of temperature and equivalent thermal dose.

This invention relates to the field of biodetection technology, specifically to a non-isolated digital multiplex detection method based on a DNA walker and TdT enzyme amplification. The method first utilizes coding microspheres with capture elements coupled to their surface to specifically bind to target miRNAs or proteins in the sample; then, specific guide DNA (gDNA) corresponding to the target molecule is released through enzymatic digestion or thermal denaturation; the gDNA then... Pf Under the action of Ago endonuclease, the DNA walker on the surface of the encoded microspheres is activated, cyclically cutting the orbital strand to generate multiple DNA fragments with 3'-hydroxyl ends. Finally, TdT enzyme is used to extend fluorescently labeled nucleotides at the hydroxyl ends, achieving in-situ signal amplification, and digital quantitative analysis is performed on the fluorescence signal of the encoded microspheres. This invention achieves high-sensitivity detection of multiple targets in a single reaction without chamber isolation, with detection limits for each target molecule reaching the fM or fg / mL level, providing an effective technical means for the early screening of major diseases such as Alzheimer's.

The invention belongs to the technical field of bioactive protein peptides / protein hydrolysates, and particularly relates to an anti-aging soybean protein peptide and a preparation method thereof. Soybean protein is used as a raw material, the soybean protein is firstly subjected to thermal denaturation treatment to change the secondary structure of the soybean protein, and then the alkaline subunit amount of the thermal denaturation soybean protein is increased by utilizing high static pressure physical treatment, so that the exposure of hydrophobic groups in soybean protein molecules and the increase of enzyme action sites are facilitated, and the proteolysis rate is further increased. And finally, carrying out enzymolysis on the treated soybean protein by using alkaline protease to obtain the soybean protein peptide. The soybean protein peptide obtained by the preparation method disclosed by the invention has very strong antioxidant activity and a good anti-aging effect.

A method for determining or verifying the identity of a protein based on a melting curve obtained from a nano-differential scanning fluorimeter (nanoDSF). The method uses a machine learning model trained on a set of proteins of known identity and includes acquiring, through use of nanoDSF, a set of data points representative of a thermal denaturation curve (melting curve) of a test protein, generating a set of features derived from the melting curve, and providing the set of features as inputs to the trained machine learning model to obtain a corresponding output indicative of an identity of the test protein. Methods for training the machine learning model are provided, as well as systems including processor-executable instructions that, when executed by a processor, cause the system to perform the method for determining or verifying protein identity.

The application discloses a preparation and separation and purification method of deer antler peptide by directional enzymolysis, and belongs to the technical field of bioactive peptide preparation. Deer antler collagen is subjected to mechanical energy auxiliary treatment in an alkaline medium under low-temperature conditions lower than the thermal denaturation temperature of the deer antler collagen, so that the triple helix structure is reversibly unfolded to increase the accessible surface area of the protease and the conformation of the active segment of the ginseng protein is retained. The treated deer antler collagen and the ginseng protein are prepared into the same substrate solution and subjected to synergistic enzymolysis. Alkaline protease, collagenase and aminopeptidase are used for directional enzymolysis in steps, so that the product is directionally enriched with two characteristic dipeptides, prolyl-hydroxyproline and hydroxyprolyl-glycine. The dipeptides are separated by three steps of ultrafiltration, gradient elution of a macroporous resin and reverse phase preparative chromatography in series, so that the low-temperature mechanical energy reversible unfolding replaces the high-temperature pre-denaturation, the contradiction between the two types of raw materials in the treatment temperature window is eliminated, the two types of raw materials can be synergistically treated in the same system, and the dipeptide yield is higher than that of the separate enzymolysis.

The invention provides an octagonal duck meat marinating parameter detection, regulation and control system, relates to the technical field of food production equipment, and solves the problem that a traditional model is difficult to consider the oily degree and the tasty depth of duck meat by constructing a two-dimensional dynamic marinating state space containing a fluid shear resistance characteristic entropy and a system reheating impedance phase angle. The fluid characteristics and the muscle fiber thermal denaturation degree are solved by utilizing motor current harmonic analysis and thermal impulse response cross-correlation analysis, so that non-intrusive accurate judgment on the marinating liquid emulsification grade and the duck meat curing stage is realized; aiming at the working condition of heat retardation, the cooperative closed-loop regulation and control module reduces the viscosity by improving the frequency of a circulating pump and utilizing the shear thinning characteristic, and limits the heating duty ratio to prevent coking, so that the limitation of optical and contact type detection under the severe working condition is overcome, the taste and flavor of marinated duck meat are ensured, and the whole-process self-adaptive closed-loop regulation and control are realized.

The invention provides an amplification library building method for high-throughput sequencing verification of synthetic genes. The amplification library building method comprises the following steps: obtaining a joint connection product of which two ends are connected with double-chain joints after a to-be-detected synthetic gene sequence, and carrying out first PCR (Polymerase Chain Reaction) amplification by adopting a first gene specific primer by taking the joint connection product as a template to obtain a first amplification product; taking the first amplification product as a template, and carrying out second PCR amplification by adopting a second gene specific primer to obtain a second amplification product; performing thermal denaturation single-chain separation and single-chain cyclization on the second amplification product to obtain single-chain circular DNA; the preparation method comprises the following steps: carrying out DNB preparation on single-stranded circular DNA to obtain DNB nanospheres; and sequencing the DNB nanospheres to obtain synthetic gene sequence information. The method verifies that the synthesized gene is high in flux, and the flux requirement of large-scale gene synthesis can be met.

The invention discloses a shark source single-domain antibody targeting LRP6 and application of the shark source single-domain antibody. The single-domain antibody has an amino acid sequence as shown in SEQ ID NO: 1, shows nanomole-level high affinity to LRP6 protein, has an affinity constant KD value of 1.8 * 10 <-9 > M, shows excellent physical and chemical stability, has a thermal denaturation temperature as high as 75.6 DEG C, and can still retain 92% or more of binding activity after being incubated for 2 hours under a strong acidic condition. Meanwhile, the antibody has high specificity, and the cross reaction rate of the antibody with homologous protein LRP5 and homologous protein LDLR is lower than 4%. The invention also provides a composition for reducing hypertension, hyperglycemia and hyperlipidemia, which contains the single-domain antibody and a specific natural plant extract composite component. Experiments show that the composition can synergistically act on a plurality of pathological links of the metabolic syndrome, the symptoms of hyperglycemia, hyperlipidemia and hypertension are remarkably improved in an animal model, the effect is better than that of a single component and a traditional control drug, and a brand new multi-target treatment scheme is provided for prevention and treatment of the metabolic syndrome.

The application relates to the technical field of preparation of canine interferon, and particularly provides a preparation method and a preparation device of canine interferon. The preparation device of the canine interferon comprises a frame body assembly, a barrel body assembly, a driving assembly, a homogenate synchronization assembly and a homogenate assembly. The application can realize controllable grading crushing, guarantee sufficient crushing of bacteria bodies, complete release of products, improve crushing efficiency, avoid excessive crushing to cause fine crushing of the bacteria bodies, effectively absorb instantaneous impact force generated by material impact, avoid excessive crushing, reduce abrasion and vibration of the equipment, provide uniform and stable temperature control for the barrel body assembly, effectively guarantee the biological activity and purity of the products. In addition, IPTG inducer is used in cooperation with low-temperature constant-temperature induction to guarantee stable and high expression of target proteins, reduce the risk of protein thermal denaturation, reduce the difficulty of inclusion body treatment, and use multi-step chromatography purification to guarantee product quality and long-term stability of the products.

The application provides a high-efficiency extraction method of animal skin enzymatic gelatin. The method is characterized in that the expansion degree and thermal denaturation law of collagen fibers of animal skin are controlled through the synergistic effect of acid, alkali, salt and heat, the exposure degree of the specific site of protease in the collagen fibers is controlled, the endo-type hydrolysis of the protease on the collagen fibers is promoted, the gelatin extraction rate and gel strength are improved, and the performance of the gelatin can meet the quality standard of industrial gelatin without desalting treatment. The method can realize one-time extraction of animal skin gelatin, the gelatin yield is more than 90%, the gel strength is more than 200 Bloom g, the production efficiency of the gelatin can be obviously improved, and good energy-saving and emission-reducing effects are achieved. The method has important significance for developing the high-efficiency preparation technology of high-performance animal skin gelatin.

This invention discloses a method for preparing platelet lysis buffer and its application, belonging to the field of cell biotechnology. The invention employs X-ray irradiation inactivation technology, irradiating plasma supernatant at a dose of 25-35 kGy under low-temperature and light-protected conditions (2-8℃) to thoroughly inactivate immunologically active substances and pathogenic microorganisms, while avoiding thermal denaturation of growth factors. This is combined with segmented ultrasonic cyclic lysis and ice-water bath temperature control (≤37℃) to ensure complete platelet lysis and reduce damage to active factors. Further treatment with calcium gluconate removes fibrin, preventing the precipitation of flocculent material. The preparation method of this invention is process-controllable and highly safe, with a core growth factor retention rate ≥80% and a fibrin removal rate exceeding 92%. The prepared platelet lysis buffer can be added to mesenchymal stem cell culture medium, effectively meeting the high-quality requirements of clinical-grade cell culture.

The invention belongs to the technical field of protein purification, and particularly relates to a purification method for expressing heat source superoxide dismutase based on escherichia coli. The invention provides an integrated process of gradient salting-out screening, dynamic thermal denaturation pretreatment and targeted chromatography, and efficient purification is realized through triple synergistic effect. According to the strategy, the traditional mindset is broken through, a gradient heating mode and ammonium sulfate salting-out optimization are combined for the first time, the activity of heat-sensitive target protein is reserved, and selective removal of impure protein is achieved.

The invention belongs to the technical field of food science and bioengineering, and particularly relates to a low-sensitization protein food formula and a preparation method thereof. The method aims at solving the problems that current single physical or enzymatic treatment is difficult to give consideration to reduction of sensitization and maintenance of nutritional functions, polysaccharide is weak in action with protein under a neutral condition, and the direct compounding effect is limited. The preparation method comprises the following steps: carrying out thermal denaturation pretreatment on soybean protein, treating the soybean protein with polyphenol, transglutaminase and pectin, then carrying out calcium ion pre-crosslinking and acid-induced gelation to form a stable gel structure, and connecting the protein and the pectin by using the polyphenol and the transglutaminase as molecular bridging agents, so as to obtain the protein-pectin composite material. The methacrylic acid copolymer and the hydroxypropyl methylcellulose phthalate are respectively used for coating protection, so that the combination efficiency and the specificity of the compound under neutral pH are remarkably improved; controllable construction of the compound is achieved through a directional assembly technology, and the sensitization of the obtained product is remarkably reduced.