15 results about "Protease digestion" patented technology

This invention discloses a method for preparing and applying plant sulfonated small peptide PSK based on synthetic biology, belonging to the field of biotechnology. Based on the design concept of synthetic biology, this invention constructs an orthogonal synthetic system composed of genetic code expansion technology and SUMO fusion protease digestion technology. A specific PSK dotted vector and a pUltra-sY vector are co-transformed into *E. coli* to construct a genetically engineered strain. The sulfonated small peptide PSK precursor is expressed in this genetically engineered strain, and mature plant sulfonated small peptide PSK is obtained through specific hydrolysis by SUMO protease. This method significantly improves the synthesis efficiency and purity of sulfonated small peptides and has broad application prospects in agricultural production and biotechnology.

This invention provides a method for forming a mammalian acoustic extracellular matrix (ECM) hydrogel without the use of acidic or alkaline solutions or protease digestion, as well as the resulting acoustic ECM hydrogel. [Solution] A method for producing an extracellular matrix (ECM) hydrogel is provided, comprising using ultrasonic frequencies to solubilize mammalian ECM in a liquid to produce a liquid-phase acoustic ECM hydrogel.

The application discloses a DNA / RNA co-extraction method and kit based on a double-water-phase temperature-induced phase separation, which utilizes polyethylene glycol-sodium citrate to form a primary double-water-phase system at 18-25 DEG C, and by adding an ionic liquid modified temperature-sensitive polymer (IL-NIPAM) with a LCST of 32 DEG C plus or minus 1 DEG C, the IL-NIPAM selectively binds with RNA, and the separation of RNA and DNA in a biological sample is realized by temperature regulation. The DNA / RNA co-extraction kit includes a double-water-phase premix, and the IL-NIPAM is added in the double-water-phase premix in advance or during use, and the use concentration of the IL-NIPAM is 0.5-1.5% w / w. The method does not need proteinase K digestion and does not need to add organic solvents, and can quickly and efficiently recover DNA, mRNA and miRNA, and is suitable for the co-extraction of nucleic acids from trace samples such as fingertip blood, dry blood spots and FFPE sections, and is suitable for automation and large-scale production.

In the analysis method according to the present invention, a predetermined peptide is separated by immunoprecipitation using an antibody which specifically binds to either an N-terminus or a C-terminus of the predetermined peptide. The separated predetermined peptide is digested with a protease to prepare peptide fragments, and among the peptide fragments, a peptide fragment at a terminus opposite to a terminus binding to the antibody is mass-spectrometrically detected.

The invention belongs to the technical field of biological tissue engineering, and discloses a fat acellular extracellular matrix hydrogel preparation method, which comprises: (1) cutting cleaned fresh fat into pieces, and repeatedly freezing and thawing with a hypertonic salt solution; (2) homogenizing the frozen and thawed fat at low temperature, and collecting and precipitating; (3) re-suspending the precipitate by using PBS, oscillating in a shaking table, centrifuging at room temperature, and collecting the precipitate again; (4) carrying out degreasing and decellularization treatment on the collected precipitate by using a composite eluent; and (5) washing the precipitate with isopropanol for 24 hours, finally repeatedly washing the precipitate with distilled water, and collecting the precipitate, namely the fat acellular matrix. And (6) digesting the fat extracellular matrix with pepsin hydrochloride, and adjusting the pH value to be neutral with sodium hydroxide to obtain the fat extracellular matrix hydrogel. According to the preparation method of the fat extracellular matrix hydrogel, biological enzyme and high-speed centrifugation are not needed in the decellularization stage, the technology is simple, time consumption is short, the yield is high, the fat extracellular matrix prepared through the method is easy to digest, and the prepared hydrogel solution is uniform, good in injectability and capable of being used for production of a filling material for injection.

Disclosed herein are polypeptides comprising an extended recombinant polypeptide (XTEN) comprised of a plurality of overlapping sequence motifs and one or more barcode fragments releasable upon protease digestion and detectable from ail other proteolytic-ally releasable fragments. Certain embodiments of these polypeptides further comprise a biologically active polypeptide, wherein advantageous embodiments thereof comprise a releasable segment capable of proteolytic cleavage that cleaves the linkage between the XTEN polypeptide and the biologically active polypeptide. Methods of making and methods of using said polypeptides are also disclosed.

The invention relates to the technical field of borosilicate glass, in particular to a sample DNA (deoxyribonucleic acid) rapid extraction method which comprises the following steps: step S1, sample pretreatment: taking 20-100 mu L of liquid sample or 25-50 mg of solid sample, adding 100-200 mu L of pretreatment liquid with the pH value of 7.5-8.0, and uniformly mixing by vortex for 10-30 seconds; s2, synergistic cracking and adsorption: adding 150-200 [mu] L of a composite cracking solution and 10-20 [mu] g of modified diatomite into the pretreatment solution, incubating at 55-60 DEG C for 3-4 min, and reversing and uniformly mixing twice during incubation; s3, impurity removal: adding 150-200 [mu] L of a washing liquid I, standing at room temperature for 1 min, and pouring to remove a supernatant; then adding 200 [mu] L of a washing liquid II, standing for 30 s, and dumping to remove the supernatant. By optimizing the composite lysate, the modified diatomite and reaction conditions, rapid DNA extraction of which the whole process is less than or equal to 8 minutes without protease K digestion and high-speed centrifugation is realized, the method has wide sample adaptability, high automation compatibility and stable DNA yield and purity, the extraction efficiency is greatly improved, and the operation cost and threshold are reduced.

Provided is a novel antibody analysis method that does not require capture or immobilization through an Fc structure. Specifically provided is a method for preparing a peptide fragment, the method comprising: a substrate immobilization step in which a target substrate antibody 125 in a biological sample is immobilized in a pore 129 of a porous body 120 that has affinity for a Fab domain of the substrate antibody 125; and a digestion step in which the porous body 120 on which the substrate antibody 125 has been immobilized is brought into contact with fine particles 110 each having a surface on which a protease 115 has been immobilized, whereby the digestion of the substrate antibody 125 with the protease is performed and a peptide fragment of the Fab domain is produced. In the substrate immobilization step, the Fab domain of the substrate antibody 125 is immobilized in the pore 129 of the porous body 120, and the average particle diameter D11 of the fine particles 110 is larger than the average pore diameter D12 of the porous body 120.

We disclose a composition comprising a plurality of plant derived peptides obtained by protease digestion prepared from a plant of the genus Brassicae and its use in the enhancement of cellular metabolism and in the treatment of diseases, for example diseases associated with muscle atrophy such as muscular dystrophy or sarcopenia.

The present invention relates to barcoded XTEN polypeptides and compositions thereof and methods of making and using the same, specifically disclosing polypeptides comprising an extended recombinant polypeptide (XTEN) comprising a plurality of overlapping sequence motifs and one or more barcode fragments that are releasable upon protease digestion and detectable from all other proteolytically releasable fragments. Certain embodiments of these polypeptides further comprise a biologically active polypeptide, with advantageous embodiments thereof comprising a proteolytic cleavage releasable segment capable of cleaving the linkage between the XTEN polypeptide and the biologically active polypeptide. Methods of making the polypeptides and methods of using the polypeptides are also disclosed.

Provided herein are methods and systems for the simultaneous targeted detection and sequencing of DNA, RNA, and Protein. In typical embodiments, the DNA, RNA, and Proteins are detected, characterized, and sequenced using just a single mammalian cell. One embodiment of detecting and characterizing DNA, RNA, or protein from a mammalian cell includes encapsulating a single cell in a drop and performing a protease digest on the encapsulated cell drop, performing a reverse transcriptase reaction; performing a droplet merger with barcoding PCR reagents and barcoding beads; performing a PCR reaction to attach the cell barcodes to the DNA targeted amplicons, RNA targeted amplicons, and protein tag amplicons, where all amplicons from the same emulsion contain the same cell barcode; and detecting and characterizing a DNA, RNA, or protein amplicon by sequencing the cell barcode incorporated into each amplicon.

Provided herein is a sensitive, quantitative, and scalable targeted proteomics assay of Alzheimer's Disease biomarkers representing neuronal, glial, vasculature and metabolic pathways. The biomarkers are protease-digested peptides selected from biological samples of individuals having normal Aβ and Tau levels (AT−) and from symptomatic and asymptomatic individuals having low Aβ and high Tau levels (AT+). The assay uses selective reaction monitoring-based mass spectrometry (SRM-MS) of peptides in the biological samples after digestion.

To provide a bispecific T cell engager (TCE) cancer therapeutic agent having an increased therapeutic index.SOLUTION: (a) an extended recombinant polypeptide (XTEN), wherein the XTEN comprises a barcode fragment (BAR) that is releasable from the polypeptide upon digestion by a protease; And (c) a polypeptide having N - and C-terminal amino acids comprising a release segment (RS) located between the XTEN and the diabody construct, wherein the diabody construct specifically binds to CD3 and comprises CDR - L1, CDR - L2 and CDR - L3, and CDR - H1, CDR - H2 and CDR - H3, wherein the CDR - H3 comprises a specific amino acid sequence, wherein the first antigen-binding fragment (AF1) specifically binds to human epidermal growth factor-receptor 2 (). HER2 AF2.SELECTED DRAWING: None

The present invention addresses the problem of providing a material for systematically exploring and identifying specific epitopes for various diseases including cancer. In addition, the present invention addresses the problem of establishing a disease analysis method using specific epitopes for various diseases including cancer. The present invention relates to a glycopeptide in which, in a peptide comprising the amino acid sequence: TTPPTTATPIR, one sugar chain comprising six or less sugars is bonded to each of one or more threonine, and the sugar chain is an O-bonded sugar chain. Furthermore, the present invention relates to a method for analyzing a glycopeptide, the method comprising analyzing whether or not a glycopeptide is contained in a glycopeptide-containing sample obtained by protease digestion of serum or plasma, the glycopeptide is obtained by bonding one sugar chain comprising six or less sugars to one or more threonine in a peptide comprising the amino acid sequence TTPPTTATPIR, and the sugar chain is an O-bonded sugar chain.

A recombinant small-molecule collagen, an expression system thereof and a preparation method thereof are provided. The recombinant small-molecule collagen derived from a plurality of sources of type-III and type-XVII collagens, and successfully expresses the recombinant small-molecule collagen. The present disclosure establishes one expression system of a small-molecule collagen and a preparation method of the small-molecule collagen, and includes an engineered exclusive chassis cell, a designed tandem repeat sequence of the small-molecule collagen, and a recombinant vector. The expression system or method of the present disclosure significantly enhances the yield of the small-molecule collagen. The present disclosure avoids the cost caused by in vitro protease digestion and the risk of exogenous protein residues, while also reducing the time and cost of a subsequent purification process.