151 results about "Peptide bond" patented technology

This invention provides a tetrahedral antibody comprising a first, second, third, and fourth domain, wherein the first and second domains are Fab or Fc domains; wherein each of the first and second domains comprise a first polypeptide chain comprising a first N-terminus of the domain, and a second polypeptide chain comprising a second N-terminus of the domain; wherein the first N-terminus of the first domain and the first N-terminus of the second domain are joined to each other by a non-peptidyl linkage, which can be a covalent linkage or a non-covalent linkage between first and second dimerizing polypeptides attached to the first N-termini of the first and second domains, respectively; and wherein the third and fourth domains are attached at their respective C-termini to the second N-termini of the first and second domains, respectively, or the N-termini of the first and second dimerizing polypeptides.

The present invention relates to a compound binding to an endogenous receptor, said compound comprising (i) an oligopeptide comprising a dipeptide with Trp being the C-terminal amino acid of said dipeptide, wherein said Trp is replaced with an α-amino acid Xaa2, whereby the stability in serum or plasma of the peptide bond connecting Xaa2 to the N-terminally adjacent amino acid is increased as compared to the peptide bond connecting Trp to the N-terminally adjacent amino acid in an otherwise identical compound; and (ii) a moiety capable of generating therapeutically effective radiation, said moiety being covalently bound to said oligopeptide.

The invention provides an amino acid skeleton ionizable lipid as well as a preparation method and application thereof, the amino acid skeleton ionizable lipid is modified by taking amino acid as a core, has more ester groups and peptide bonds, and can be quickly hydrolyzed by enzyme after RNA is effectively released in vivo; the transfection body is provided with four tail structures, the cross sectional area of the lipid tail can be increased, drugs such as RNA are helped to escape from an endosome, and then the transfection effect is enhanced; the charge capable of ionizing the lipid is electrically neutral under physiological conditions, so that the cytotoxicity caused by excessive positive charges is reduced, the stability of the lipid nanoparticles is further improved, the cycle time of the loaded nucleic acid medicine is prolonged, and the pharmacokinetic characteristics are improved. The LNP prepared from the ionizable lipid, auxiliary phospholipid, cholesterol and PEG lipid provided by the invention has more excellent nucleic acid carrier performance, and can effectively deliver nucleic acid drugs such as siRNA, mRNA, pDNA and the like into cells to play a role.

Compositions and related methods and uses are provided comprising sotebercept and variants thereof wherein the variants undergo a peptide bond cleavage between arginine 4 and serine 5 (Arg (4)-Ser (5) of the amino acid sequence (SEQ ID NO: 2) of one or two monomeric peptide chains of sotebercept, calculated based on peptide mass fingerprinting analysis, and wherein the variants have a peptide bond cleavage between arginine 4 and serine 5 (Arg (4)-Ser (5) of one or two monomeric peptide chains of sotebercept. The variant content is less than or equal to about 10%. The invention further provides a pharmaceutical preparation containing the composition and a preparation method of the pharmaceutical preparation. In addition, the invention also provides a detection method of the variant and application of the variant in quality inspection or quality control of a product containing sotebercept.

The invention relates to molecular glue for forming fimbriae related protein based on SpaA isopeptide bonds of Vinci bacteria. The invention also relates to a nanoparticle vaccine composition formed by the molecular glue, and the immunogen is covalently displayed on the surface of the nanoparticle by the molecular glue.

The invention belongs to the technical field of enzyme engineering and gene engineering, and particularly relates to a recombinant immobilized enzyme, an expression gene thereof and application of the recombinant immobilized enzyme in production of macrocyclodextrin. According to the preparation method, SpyTag and SpyCatcher peptides are fused to the N end of isoamylase and the N end of 4-alpha-glucanotransferase respectively, the 4-alpha-glucanotransferase modified by the SpyTag and the isoamylase modified by the SpyCatcher are obtained, and after spontaneous isopeptide bonds are formed between the SpyTag and the SpyCatcher, the self-assembled macrocyclodextrin production enzyme cluster is obtained. The obtained macrocyclodextrin production enzyme cluster forms an enzyme aggregate with a three-dimensional structure under the traditional glutaraldehyde-mediated catalytic action, namely the recombinant immobilized enzyme. The recombinant immobilized enzyme disclosed by the invention can be repeatedly utilized, is simple and convenient to operate, is easy to separate a converted product, and can be applied to industrial production of the macrocyclodextrin.

The present invention pertains to specific bioparticules at the surface of which are expressed multimeric protein. The bioparticles according to the present invention comprise an envelope consisting of a plasma membrane; and at least one type I or II transmembrane fusion protein anchored in said membrane, said fusion protein comprising successively a) a first monomer of a multimeric protein of interest, b) a coiled-coil domain or oligomerization sequence; and c) a domain for anchoring in the plasma membrane, consisting of a transmembrane segment and a cytosolic segment. Fragments a) and b) are exposed at the surface of the bioparticle, and fragment a) is bound to a second monomer of said multimeric protein by means of a bond which is not a peptide bond. The bioparticles according to the present invention can be used in therapy such as immunotherapy. The present invention also pertains to methods for producing such bioparticles.

The invention discloses a sea-buckthorn small molecule peptide and a preparation method thereof, and relates to the field of biological medicine and food processing. According to the method, sea-buckthorn pulp residues and sea-buckthorn seed residues are used as composite raw materials and are mixed and stirred to form homogenate, and then cell wall structures are destroyed through ultrasonic wall breaking treatment to release protein components; then cutting off a peptide bond under the action of compound enzymes (plant protease, neutral protease and pepsase) by adopting an ultrasonic-enzyme synergistic hydrolysis process to generate a small molecule peptide fragment; after removing impurities through centrifugal separation, purifying through an ultrafiltration membrane (the molecular weight cut-off is 1-3kDa) to obtain a small molecule peptide solution; and performing debitterizing treatment by using macroporous adsorption resin, and preparing sea-buckthorn small molecule peptide powder by using a vacuum freeze-drying technology. According to the method, through ultrasonic-enzyme synergistic interaction, multi-stage purification combination and low-temperature keep-alive processes, the yield, molecular weight uniformity and antioxidant activity of small molecular peptides are remarkably improved, and high-valued utilization of sea-buckthorn processing by-products is achieved.

The invention discloses an in-vitro biochemical test method for evaluating the anti-wrinkle effect of cosmetics and raw materials. The in-vitro biochemical test method comprises the following steps: step 1, preparing a working solution; 2, preparing a sample solution; step 3, adding an enzyme working solution; step 4, loading samples in groups; 5, adding a substrate and reacting; step 6, developing treatment; step 7, determining absorbance; 8, calculating, analyzing and processing data; and step 9, result judgment: analyzing and evaluating the anti-wrinkle effect of the sample through a statistical means. According to the method, based on a collagenase inhibition test, when peptide bonds between GLY and GLY in Z-GLY-PRO-GLY-GLY-PRO-ALA-OH are specifically hydrolyzed by collagenase, GLY-PRO-ALA is released, and the activity of the collagenase is indirectly evaluated by measuring the change of a light absorption value at 565 nm after the GLY-PRO-ALA reacts with a color developing agent, so that whether a sample to be detected has an anti-wrinkle effect or not is judged. The method is economical and fast, the test process is controllable, the test system is standardized, and result deviation caused by individual difference can be avoided.

This invention discloses methanogenic lyases with the same protein family annotation as PeiR lyases and their applications. These enzymes are obtained through homology screening based on the known characteristics of PeiR lyases targeting peptide bonds in the methanogenic cell wall, using the Pfam protein family database annotation. PeiR lyase proteins participate in the biological process of hydrolyzing archaea cell walls, effectively killing methanogens and reducing methane production. Proteins homologous to this protein have potential methane-reducing potential. This invention integrates rumen microbial metagenomic sequencing data, uses a series of bioinformatics software to screen methanogenic viral proteins, and combines Pfam functional domain annotation for homology analysis, ultimately identifying a series of lyases. These lyases were successfully expressed in a prokaryotic expression system, and in vitro gas production experiments also showed that the crude enzyme solution significantly reduced methane production.

This invention belongs to the field of photocrosslinking probe technology, specifically relating to a GUSB photocrosslinking probe, its preparation method, and its application. The GUSB photocrosslinking probe has the general chemical formula C0. 13 H 18 N2O6-R, wherein R is any one of the oxalic group elements or peptide bonds. The R group covalently connects the glucuronic acid recognition core and the photosensitive unit in the GUSB photocrosslinking probe, and the R group contains no more than 4 atoms, resulting in a small size and low steric hindrance. The GUSB photocrosslinking probe provided by this invention uses glucuronic acid as the recognition core, combined with a small-volume diazacyclopropane photosensitive unit and a terminal alkyne group, to achieve µM-level in-situ covalent capture of active GUSB in complex systems such as cells or tissues. The probe of this invention significantly improves positioning accuracy, sensitivity, data availability, quantification, and enrichment, while also possessing advantages such as modular synthesis, mild conditions, and strong system compatibility, effectively addressing the technical shortcomings of existing technologies that struggle to accurately obtain GUSB activity in situ under low-dose conditions.

This invention discloses a methanogenic lyase homologous to PeiR lyase and its applications. The PeiR lyase protein, targeting the peptide bonds of the methanogenic cell wall, participates in the biological process of hydrolyzing the archaea cell wall, effectively killing methanogens and reducing methane production. Proteins homologous to this lyase possess potential methane-reducing potential. This invention utilizes a series of bioinformatics software to identify protein sequences encoded by methanogenic viruses from a large amount of rumen microbial virome sequencing data, and analyzes the structural homology between these proteins and PeiR. Ultimately, this invention successfully obtained a series of methanogenic lyases homologous to PeiR. To achieve the expression of these lyases, this invention designed multiple primers to synthesize the target sequences and successfully expressed them in a prokaryotic expression system. In vitro gas production experiments with the crude enzyme solution showed that these lyases are effective in reducing methane production.

The invention discloses a fatty cation amphiphilic peptide simulant as well as a preparation method and application thereof. The preparation method comprises the following steps: firstly, developing an amphipathic cationic peptide simulant system based on cysteine, then introducing ethylenediamine as a cationic group by forming a peptide bond at the terminal of carboxyl, and introducing a hydrophobic group containing an aromatic ring to a sulfydryl side chain; wherein the cationic group provides positive charges and hydrophilicity and is beneficial to electrostatic interaction with a bacterial membrane with negative charges, and the hydrophobic group is beneficial to insertion of the membrane into a phospholipid bilayer; in addition, the N terminal is modified by fatty acids with different chain lengths, the amphiphilic balance of the cysteine-based cationic peptide simulant is finely adjusted, and finally the fatty cationic amphiphilic peptide simulant is synthesized. The peptidomimetic has good antibacterial activity, is not easy to induce bacteria to generate drug resistance, and can be used for preparing drugs for treating, improving and / or preventing infection of drug-resistant bacteria and / or drug-resistant fungi.

The present invention relates to vaccine compositions, most notably vaccine compositions wherein the antigenic component is large, for example over 50 kDa, or multimeric, i.e. comprised of subunits. Such antigenic components are of particular interest, because they may represent antigenic components from pathogens that currently it is not possible to vaccinate against. The invention relates to a composition comprising a particle displaying an antigenic component, wherein said composition comprises an antigenic component comprising a first peptide tag, and a moiety comprising a second peptide tag, wherein the antigenic component and the moiety are linked via an isopeptide bond between said first and second peptide tags, and wherein the antigenic component is over 50 kDa, or alternatively is multimeric.

The present invention relates to a method for modifying meat, including bringing the meat in contact with a Bacillus amyloliquefaciens-derived neutral protease having the following properties (a) to (e), and thermolysin; (a) being an enzyme that hydrolyzes a substrate having a peptide bond to release the peptide, (b) having an optimal pH of 7, (c) being stable at pH=5 to 7, (d) having an optimal temperature of 50°C to 55°C, (e) being a metal protease inhibited by ethylenediaminetetraacetic acid (EDTA), and a composition for modification, containing said neutral protease and thermolysin. According to the present invention, meat can be tenderized conveniently in a short time, and ease of loosening can be imparted to the meat.

This invention provides a method for producing Clostridium neurotoxins containing an exogenous activation loop, modified Clostridium neurotoxins, and pharmaceutical compositions. [Solution] A method for proteolytically processing a single-chain Clostridium neurotoxin into a corresponding double-chain Clostridium neurotoxin, comprising: providing a single-chain Clostridium neurotoxin; and contacting the single-chain Clostridium neurotoxin with enterokinase or factor Xa, wherein the single-chain Clostridium neurotoxin has the polypeptide sequence Cys-(Xaa) a -Ile-Asp / Glu-Gly-Arg-(Yaa) b - A method in which an activation loop containing Cys (SEQ ID NO: 1) is present, a=1 to 10, b=4 to 15, and enterokinase or factor Xa hydrolyzes the peptide bond of the activation loop, thereby producing a double-stranded Clostridium neurotoxin.

The invention discloses a synthesis method of ubiquitin (1-74) NHNH2, and belongs to the technical field of protein synthesis. The synthesis method comprises the following steps: utilizing a prokaryotic expression system, expressing a full-length protein of Ub (1-76) and a Josephin protein coded by Burkholderia pyrrocinia by virtue of an escherichia coli BL21 (DE3) strain, hydrolyzing the Ub (1-76) by virtue of the obtained 6His-BpJOS, cutting a peptide bond between Ub Arg-74 and Gly-75, and capturing a generated thiolipid intermediate by virtue of hydrazine hydrate, so as to obtain the Ub (1-74) NHNH2. According to the present invention, the ubiquitin cleavage enzyme BpJOS catalyzed in-situ activation process is adopted, such that the preparation yield is high, the operation is simple, the large-scale preparation can be achieved, the synthesis cost is low, and the method can be used for synthesizing the multi-ubiquitination modified fragment and synthesizing the Ub (1-74) probe as the precursor, and has wide application prospects.

The present invention relates to a polypeptide forming one part of a two-part linker wherein the polypeptide (protein) spontaneously forms an isopeptide bond with a peptide tag, i.e. The other part of the two-part linker. The invention also relates to a method for producing a polypeptide of the invention and to the use of said polypeptide. According to the present invention, the reaction rate of the Sdy-Catcher variant obtained through mutation is significantly improved compared to the reaction rate of Sdy-Tag, such that the ideal direction is provided for the searching of polypeptides with characteristics of high binding rate and good binding effect in the field.

The invention relates to a molecular peptide mutant, the amino acid sequence of which is as shown in SEQ ID NO: 1. In the invention, SpyCatcher is designed and modified to obtain a molecular peptide SpyCatcher-21 with stimulus response to pH on the basis of not affecting the formation of isopeptide bonds, and Pro is introduced into a key loop of the SpyCatcher-21 through analysis of the crystal structure to reduce the flexibility of the loop and obtain a mutant SpyCatcher-21_A82P, which can raise ligation efficiency with SpyTag. The SpyCatcher-21_A82P can be used to achieve double-enzyme catalysis according to objective needs by changing the pH of the environment to obtain different degrees of coupling, or obtain a three-enzyme coupled catalytic system through electrostatic interaction with a positively charged enzyme.

The present invention relates to a hyaluronic acid-based hydrogel which is a hyaluronic acid-peptide crosslinked body crosslinked using a peptide crosslinking agent. More specifically, the present invention relates to a hyaluronic acid-based hydrogel and a method for producing same, wherein a crosslinked body having novel physical properties is obtained using a relatively small amount of a crosslinking agent that forms peptide bonds, unlike conventional crosslinking agents, and the hyaluronic acid-based hydrogel has the advantages of: being safe and having few side effects; the physical properties of a filler being adjustable according to the amount of peptide crosslinking; and having a high elasticity ratio.

The invention discloses a SpaD-based polypeptide-protein reaction pair, and belongs to the technical field of protein engineering. The D2-D3 structural domain of the corynebacterium diphtheriae pilus protein SpaD is split into reaction pairs in two modes, the two parts can be efficiently recombined and react to generate isopeptide bonds, and covalent coupling is achieved. Wherein when the D2 structural domain is split into reaction pairs, a D214-T455 residue of an original sequence is a capturer protein (D2C), and an E182-K213 residue of the original sequence is a tag polypeptide (D2T); when the D3 structural domain is split into reaction pairs, the E182-D434 residue of the original sequence is a capturer protein (D3C), and the G438-T455 residue of the original sequence is a tag polypeptide (D3T). The reactivity of a reaction pair obtained by splitting D3 is equivalent to that of a common spy reaction pair, is not influenced by the position of a tag sequence, is orthogonal to the spy reaction pair and a probe reaction pair, is a novel protein coupling tool, and has a relatively high practical value in protein engineering.

The present disclosure provides a heterodimeric antibody, or antigen-binding fragment thereof, comprising two heavy chains with different amino acid sequences and a binding polypeptide. The heterodimer antibody, or the antigen-binding fragment thereof, include a first heavy chain comprising none or one or more modifications and the second heavy chain comprises one or more modifications and is fused to a binding polypeptide, which can form a isopeptide bond with a binding peptide. The present disclosure further provides systems and methods for purifying such heterodimeric antibodies, and antigen-binding fragments thereof.

The present invention provides: a method for determining the amino acid sequence of a peptide without depending on a database; and a derivatizing agent useful for the method. The present invention is a method for determining the amino acid sequence of a peptide, the method involving: a step for performing multiple-stage tandem mass spectrometry (MSn) on a derivatized peptide in which a derivatizing agent having a coumarin skeleton is bound to a peptide to obtain a fragment ion mass spectrum of the derivatized peptide; and a step for determining the amino acid sequence of the peptide on the basis of peaks of fragment ions of the derivatized peptide in which peptide bonds are sequentially cleaved.

The application belongs to the technical field of cosmetics and skin external preparation, and discloses a retinoid, a preparation method and application thereof. The compound is formed by covalently coupling a Schiff base or a kinin releasing enzyme cleavable peptide bond (L) and a free L-amino acid, a di- to hexapeptide or a C6-C22 fatty acid ester (P) of the retinol parent (R), and the structural general formula is R-L-P. The compound has a 12-week stability rate of ≥80% in a 25℃ aqueous preparation, is recognized by a keratin layer carboxylic acid esterase and a kinin releasing enzyme KLK-5 / KLK-7 after being applied to the skin, is subjected to enzymatic cutting in a skin acidic cuticle (pH 4.5-5.5) microenvironment, and releases free retinol at a fixed point. Compared with retinol palmitate, the in-situ release efficiency of the compound of the application is increased by 2.5-4.5 times, while the risk of systemic absorption is significantly reduced, and the incidence of sensitive skin irritation is ≤6.7%.

This invention discloses a method for rapidly constructing a complex of cholera toxin B subunit (CTB) and antigen. The vaccine complex includes a CTB-SpyTag fusion protein and at least one SpyCatcher-antigen fusion protein. It also includes a preparation method for rapidly preparing a CTB5-antigen complex by mixing CTB-SpyTag with the SpyCatcher-antigen. This invention directs the antigen to the side of the CTB pentamer that does not participate in GM1 binding, avoiding steric hindrance interference. It utilizes the SpyTag / SpyCatcher covalent isopeptide bond to obtain highly stable antigen display, reducing antigen shedding during subsequent preparation and storage. It also possesses mucosal targeting and multivalent presentation functions, providing a universal platform technology for rapidly constructing and validating broad-spectrum, combined, or personalized mucosal vaccines.

The present invention provides a compound having the structure:A-B-----Zwherein A is a biologically active structure of the compound;wherein Z is a protein component of the compound, which protein component comprises one or more polypeptides, wherein at least one of the one or more polypeptides comprises consecutive amino acids which (i) are identical to a stretch of consecutive amino acids present in a chain of an Fc domain of an antibody; (ii) bind to an Fc receptor; and (iii) have at their N-terminus a sequence selected from the group consisting of a cysteine or selenocysteine; wherein the dashed line between B and Z represents a peptidyl linkage; and wherein the solid line between A and B represents a nonpeptidyl linkage, as well as intermediates dimers thereof, and processes of producing the compounds of the invention.

To provide a cytoplasm delivery agent containing peptide and a cationic molecule used for delivering a target substance to a cytoplasm, and use thereof.SOLUTION: A cytoplasm delivery agent contains peptide including an amino acid sequence represented by the following formula (1) or a salt thereof, and a cationic molecule. X1X2X3X4 formula (1) X1 denotes an artificial amino acid residue or an aspartic acid residue in which artificial amino acid represented by the following general formula (A-1) or the following general formula (A-2) is bonded by peptide bond, X2, X3 and X4 denote alanine, leucine, isoleucine, valine or phenylalanine. In the formula, R11 and R21 denote a hydrocarbon group of carbon atom number 3 to 10 which may have a substituent group. R12 and R22 denote a protective group or a hydrogen atom.SELECTED DRAWING: None

There is provided peptidic compounds of Formula I, A or B(Rradn6-[linker]-RL-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-ψ-Xaa9-NH2). Xaa1 is D-Phe, Cpa, D-Cpa, Nal, D-Nal, 2-Nal, or D-2-Nal; Xaa2 is Asn, Gln, Hse, Cit or His. Xaa3 is Trp, Bta, Trp(Me), Trp(7-Me), Trp(6-Me), Trp(5-Me), Trp(4-Me), Trp(2-Me), Trp(7-F), Trp(6-F), Trp(5-F), Trp(4-F), Trp(5-OH), or αMe-Trp. Xaa4 is Ala or Ser. Xaa5 is Val, Cpg, or Tle. Xaa6 is Gly, NMe-Gly, or D-Ala. Xaa7 is His or NMe-His. Xaa8 is Leu or Phe. Xaa9-NH2 is a C-terminally amidated amino acid residue selected from Pro, 4-oxa-L-Pro, Me2Thz, or Thz. ψ represents a peptide bond or reduced peptide bond joining Xaa8 to Xaa9. Rradn6 is 1-5 radiolabeling groups. There is also provided the use of such compounds as imaging agents or therapeutic agents.

This invention provides high sensitivity 19 A fluorine-containing molecule suitable as an F-MRI contrast agent, and a material containing the fluorine-containing molecule as an active ingredient. 19 We provide F-MRI contrast agents. [Solution] A peptide comprising two or more amino acids linked by peptide bonds, wherein the peptide contains a fluorine-containing alkyl group and an organic radical within the same amino acid residue; the peptide according to the above, wherein the fluorine-containing alkyl group is a perfluoroalkyl group having 1 to 6 carbon atoms; the peptide according to the above, wherein the fluorine-containing alkyl group is a trifluoromethyl group or a nonafluoro-tert-butyl group; the peptide according to the above, wherein the organic radical is an N-oxyl radical: A pharmaceutical composition containing any of the above peptides; and a pharmaceutical composition containing any of the above peptides as an active ingredient. 19 F-MRI contrast agent.

The application provides a recombinant polymyxin enzyme and a preparation and application method thereof, an amino acid sequence of the recombinant polymyxin enzyme is shown as SEQ ID NO. 1, and the recombinant polymyxin enzyme is obtained through site-directed mutation on an amino acid sequence of a wild-type polymyxin enzyme derived from Brevibacillus laterosporus. The recombinant polymyxin enzyme can specifically recognize and cut a peptide bond between a tripeptide side chain and a cyclic heptapeptide ring in a polymyxin structure and a peptide bond between threonine and diaminobutyric acid inside the cyclic heptapeptide ring, and further can specifically degrade or neutralize the recombinant polymyxin enzyme of polymyxin.