88 results about "Single chain" patented technology

The application discloses an oligonucleotide hydrogel and a preparation method and application thereof, and comprises the following steps: sequentially adding oligonucleotide and PBS buffer into a metal salt solution to obtain a mixed solution; placing the mixed solution in a shaking table and oscillating and incubating; after incubation, centrifuging, discarding the supernatant and reserving the precipitate; resuspending the precipitate by adding deionized water, centrifuging, discarding the supernatant and washing to obtain the hydrogel; the application only needs single-stranded oligonucleotide, has no special requirements on the sequence composition and structure of the oligonucleotide, does not need special design, can be assembled with divalent and trivalent metal ions, has a simple process, is easy to operate, and can wrap biological enzymes in the hydrogel, realizes enzyme immobilization and recycling.

The application provides an anti-Spike protein antibody of a novel coronavirus and an application thereof. By means of genetic engineering and phage surface display library technology, specific antibodies against the S protein of the novel coronavirus are screened from a single-chain antibody library of non-immune full human sequences. The antibodies have an affinity to the S protein of the virus between 1nM and 50nM, the antibodies have an inhibiting effect on the combination of the S protein of the novel coronavirus and human receptor ACE2, and the anti-S protein antibodies of the application have a good ability to combine with the S protein and a potential neutralizing inhibiting effect. The application provides specific antibody candidate molecules for the development of diagnostic reagents, preventive and therapeutic antibody drugs against the novel coronavirus (2019-nCoV) and the treatment of other diseases such as pneumonia caused by the coronavirus.

This invention discloses direct amplification qPCR reagents, kits, and applications for whole blood samples. The amplification reagents include: a composite hot-start DNA polymerase system, a buffer system, a surfactant, an anti-contamination system, and an anti-inhibition enhancer system. The composite hot-start DNA polymerase system is obtained by mixing antibody-modified hot-start Taq enzyme and antibody-modified Taq1C2 mutant Taq enzyme in a predetermined ratio. The buffer system includes Tris-HCl, MgCl2, (NH4)2SO4, and KCl. The surfactants include Tween-20 and NP-40. The anti-contamination system includes UDG enzyme and dNTPs, wherein the dNTPs are composed of dATP, dGTP, dCTP, dTTP, and dUTP. The anti-inhibition enhancer system includes D-trehalose, L-carnitine, DMSO, BSA, and SSB single-chain binding protein.

The present application relates to a kind of based on logic gate double-target point chimeric antigen receptor modified TIL cell, lentivirus expression vector and application, belong to tumor immunotherapy and gene editing technical field.The TIL cell based on logic gate double-target point chimeric antigen receptor modified in the application, double-target point chimeric antigen receptor includes chimeric antigen receptor EGFR and chimeric antigen receptor GD2;Chimeric antigen receptor EGFR is composed of CD8 alpha signal peptide, anti-EGFR single-chain antibody, CD8 alpha transmembrane region, 4-1BB costimulatory domain and CD3 zeta intracellular signal domain in series;Chimeric antigen receptor GD2 is composed of CD8 alpha signal peptide, anti-GD2 single-chain antibody, CD28 transmembrane region, CD27 costimulatory domain and CD3 zeta intracellular signal domain in series.The present application solves the defects that lentivirus transduction targeting is poor in prior art, CAR signal activation specificity is insufficient, TIL cell is easily exhausted, has the advantages that gene integration is accurate, signal transduction is controllable, in-vivo survival time is long, can be efficiently used for the immunotherapy of double-antigen co-expression solid tumor.

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.

Provided herein are Mycobacterium smegmatis porin nanopores, systems that comprise these nanopores, and methods of using and making these nanopores. Such nanopores may be wild-type MspA porins, mutant MspA porins, wild-type MspA paralog porins, wild-type MspA homolog porins, mutant MspA paralog porins, mutant MspA homolog porins, or single-chain Msp porins. Also provided are bacterial strains capable of inducible Msp porin expression.

The application provides an anti-Spike protein antibody of a novel coronavirus and an application thereof. By means of genetic engineering and phage surface display library technology, specific antibodies against the S protein of the novel coronavirus are screened from a single-chain antibody library of non-immune full human sequences. The antibodies have an affinity to the S protein of the virus between 1nM and 50nM, the antibodies have an inhibiting effect on the combination of the S protein of the novel coronavirus and human receptor ACE2, and the anti-S protein antibodies of the application have a good ability to combine with the S protein and a potential neutralizing inhibiting effect. The application provides specific antibody candidate molecules for the development of diagnostic reagents, preventive and therapeutic antibody drugs against the novel coronavirus (2019-nCoV) and the treatment of other diseases such as pneumonia caused by the coronavirus.

The application discloses a kind of bispecific anti-CD3 / CD20 antibodies and its application in B cell lymphoma, belong to the field of biological medicine technology.The bispecific antibody uses "tandem type" molecular structure, by the single-chain antibody (scFv) of anti-CD20 and the nanometer antibody (VHH) of anti-CD3 are fused by flexible connecting peptide.The high-purity bispecific antibody BsAb-20S3V is obtained by CHO cell stable expression system, and it has nanomolar level affinity to CD20+ and CD3+ cells.Experiments in vitro prove that the antibody can effectively mediate T cell specific killing to Raji cell, and the maximum killing rate can reach 75%-90%.In vivo pharmacodynamic evaluation shows that in tumor-bearing mouse model, high-dose group can significantly inhibit tumor growth and induce tumor negative growth.The antibody has good potential to prepare B cell lymphoma treatment drugs.

This invention discloses a mutant, gene, and recombinant bacteria of a highly thermostable single-chain sweet protein. The mutant is characterized by the following amino acid sequences shown in SEQ ID NO.1: isoleucine (I) at position 5 is mutated to glutamic acid (E), glutamic acid (E) at position 23 is mutated to alanine (A), isoleucine (I) at position 26 is mutated to arginine (R), cysteine ​​(C) at position 41 is mutated to alanine (A), tyrosine (Y) at position 65 is mutated to isoleucine (I), glycine (G) at position 83 is mutated to arginine (R), and asparagine (N) at position 90 is mutated to glutamic acid (E). Based on structural analysis and computationally assisted design for protein stability, this invention achieves site-directed mutagenesis of the single-chain sweet protein, resulting in a highly thermostable mutant. Compared to the original single-chain sweet protein, the thermostable mutant of this invention exhibits higher T... m It was heated by more than 20.4°C while maintaining its sweetness.

The present disclosure provides a peptide-major histocompatibility complex (pMHC) polypeptide comprising a peptide epitope and class I MHC polypeptides. The present disclosure provides fusion molecules comprising a pMHC polypeptide and a heterologous fusion partner. The present disclosure provides single-chain T-cell modulatory polypeptides that comprise a pMHC polypeptide, one or more immunomodulatory polypeptide, and an immunoglobulin (Ig) Fc or a non-Ig scaffold. A TMP is useful for modulating the activity of a T cell, and for modulating an immune response in an individual.

The application discloses IV type recombinant collagen protein and a preparation method and application thereof, and relates to the technical field of genetic engineering. The recombinant collagen protein comprises a recombinant collagen single chain; wherein the recombinant collagen single chain is obtained by repeating a collagen monomer fragment as shown in SEQ ID NO. 2 for 2-10 times. The application designs and obtains a recombinant collagen amino acid sequence which can be stably and efficiently expressed in Pichia pastoris and has excellent biological activity by accurately screening and optimizing the functional region sequence of human natural IV type collagen. The sequence is 100% homologous to the corresponding region of human natural IV type collagen, thereby ensuring low immunogenicity of the product and excellent biocompatibility with the human body from the source.

A manufacturing method of a horizontal rotary edge single chain scraper conveyor interval tooth wear-resistant sprocket, comprising the following steps: designing and calculating various important parameters of the sprocket; machining the sprocket body according to the obtained various important parameters of the sprocket; machining the movable lining plate; assembling and fixing the machined sprocket body and the movable lining plate through bolts; and finishing and heat treating the assembled sprocket assembly. The interval tooth wear-resistant sprocket manufactured by the method has a pitch at the sprocket chain nest which is 3 times the pitch of the round link chain, the interval tooth sprocket can be matched with the conventional round link chain, a variable pitch chain does not need to be designed separately, and the universality of the interval tooth sprocket is improved; the sprocket can provide a larger space to avoid interference with the guide scraper; the upper and lower tooth chain nests of the interval tooth sprocket and the upper surface of the lower tooth are subjected to local induction quenching treatment, and the movable lining plate is arranged on the upper surface of the lower tooth to enhance the wear resistance. The application also provides a horizontal rotary edge single chain scraper conveyor interval tooth wear-resistant sprocket.

The present invention relates to: a biosensor for simultaneous multiplex detection of target nucleic acid molecules, the biosensor comprising gold nanorods with immobilized ssDNA and ssRNA; and a composition for diagnosing cancer using the biosensor. The present invention enables rapid simultaneous multiplex detection of cancer-associated cfDNA and miRNA, which are target nucleic acid molecules, to be performed with high accuracy and sensitivity, and thus can be advantageously used in the field of cancer diagnosis.

The present invention relates to the field of cell therapy. Disclosed are a fibroblast activation protein-targeting CAR T-cell, a preparation method therefor, and an application thereof. A CAR T-cell can be prepared by introducing an FAP-targeting chimeric antigen receptor (CAR) into a T lymphocyte, wherein the CAR in the CAR T-cell comprises a signal peptide, an antigen-binding domain, a hinge region, a transmembrane domain, a co-stimulatory signaling region, and a CD3 signaling domain. The CAR T-cell specifically recognizes the FAP by means of a single-chain variable fragment (scFv), which activates an intracellular signaling pathway, releasing cytokine IFN-γ, and exhibiting a cytotoxic effect on FAP+ cells, thereby achieving specific depletion of FAP+ cells at a lesion site. The CAR-T cell can be used for treating diseases characterized by upregulated FAP expression, such as fibrosis (pulmonary fibrosis, hepatic fibrosis, cardiac fibrosis, renal fibrosis, etc.), arthritis, autoimmune disorders (Crohn's disease, rheumatoid arthritis, etc.), and cardiovascular diseases, and demonstrates tremendous application potential and commercial value.

The application discloses a mutant and gene of single-chain sweet protein with high thermal stability, wherein the mutant is obtained by mutating isoleucine (I) at the 5th position, glutamic acid (E) at the 23rd position, isoleucine (I) at the 26th position, tyrosine (Y) at the 65th position, glycine (G) at the 83rd position and asparagine (N) at the 90th position in the amino acid sequence shown in SEQ ID NO. 1 to alanine (A), arginine (R), isoleucine (I), glutamic acid (E) and aspartic acid (D) respectively; the application is based on structural analysis and protein stability calculation aided design, and the single-chain sweet protein is subjected to site-directed mutagenesis, so that the mutant of single-chain sweet protein with high thermal stability is obtained; compared with the single-chain sweet protein, the mutant of single-chain sweet protein with high thermal stability has a T m increase of 18.4 DEG C or more and maintains sweetness.

The application discloses a primer group, a kit and application for screening autism spectrum disorder patients, and belongs to the field of disease diagnosis. The technical problem solved by the application is how to screen autism spectrum disorder patients in Chinese Han population. The primer group for screening autism spectrum disorder patients disclosed by the application is directed to 132 autism susceptible gene sites, and is composed of 264 single-stranded DNAs shown in SEQ ID No. 1 to SEQ ID No. 264. According to the 132 autism susceptible gene sites, a data model is established by SVM, autism spectrum disorder patients can be screened, and the data model has good accuracy, precision, recall rate, F1 score and AUC value, can provide stable and efficient prediction performance between balanced precision and recall rate, and has strong ability in distinguishing autism spectrum disorder patients and healthy individuals.

The present disclosure provides a model training method, an antibody evaluation method and an electronic device. The model training method comprises the following steps: a first training stage: training a basic model using antibody heavy chain data and antibody light chain data respectively to learn the intra-chain characteristics of antibody single chains, and obtaining corresponding heavy chain encoding parameters and light chain encoding parameters; a second training stage: taking the heavy chain encoding parameters as the initialization parameters of a first sub-model in a first interaction model and taking the light chain encoding parameters as the initialization parameters of a second sub-model in the first interaction model; training the first interaction model using heavy chain-light chain pairing data; and a third training stage: inputting the output of the first sub-model and antigen data into a third sub-model after fusion, inputting the output of the second sub-model and antigen data into a fourth sub-model after fusion, and learning the interaction characteristics between antigens and antibodies. The present disclosure trains the model in a staged manner and can output high-quality interaction characteristics between antigens and antibodies.

The present application belongs to the technical field of cellular immunization engineering, and particularly relates to a polypeptide specifically recognizing PSCA and application thereof. The polypeptide provided by the present application specifically recognizes PSCA and has good affinity performance. The polypeptide specifically targeting PSCA provided by the present application can mark PSCA positive cells, combine with PSCA antigen and PSCA positive cells, and can be used as a detection reagent for PSCA molecules and an in-vivo tracing labeling reagent. The polypeptide specifically targeting PSCA provided by the present application can also be used in an extracellular antigen recognition region of a chimeric antigen receptor, and can be used in an immune cell treatment drug targeting PSCA to kill malignant tumors expressing PSCA. The polypeptide specifically recognizing PSCA of the present application is derived from a natural full human antibody, and compared with a mouse-derived antibody, a chimeric antibody and a humanized antibody, the immunogenicity is greatly reduced, and the safety can be maximally ensured in clinical application.

The present application relates to the technical field of biological medicine, and more particularly to a multifunctional antibody K1932 binding human CD19, CD3 and Fc gamma R, wherein the multifunctional antibody is composed of a bivalent Fab fragment specifically recognizing CD19 on the surface of B lymphocyte, a bivalent single-chain antibody recognizing CD3 molecule on the surface of T lymphocyte, and a human immunoglobulin Fc domain binding with Fc receptor, wherein the single-chain antibody recognizing CD3 molecule is connected with the C-terminal of Fab light chain through a connecting peptide Linker1, and the heavy chain and the light chain of the CD3 single-chain antibody are connected by a flexible Linker2; the biological macromolecule with the three functional domains is a highly directional immunotherapy drug, which is suitable for injection administration, can guide T cells to attack CD19-positive B lymphocytes, and the Fc domain is combined with Fc gamma R, so that the in-vivo half-life of the biological macromolecule is adjusted and the unnecessary influence on normal cells is reduced.

This application discloses antibodies or antigen-binding fragments thereof that specifically bind to estradiol, immune complexes comprising the same, single-chain antibodies that specifically bind to the immune complexes, methods for their preparation, and uses. The antibody or antigen-binding fragment specifically binding to estradiol comprises a first heavy chain variable region and a first light chain variable region; wherein the first heavy chain variable region comprises first heavy chain CDR1, first heavy chain CDR2, and first heavy chain CDR3, and the first light chain variable region comprises first light chain CDR1, first light chain CDR2, and first light chain CDR3, and each of the six CDRs comprises an amino acid fragment with the sequence shown in SEQ ID NO: 1-6. Applying this antibody or antigen-binding fragment specifically binding to estradiol, along with ScFv selected based on it, to the sandwich method for detecting E2 in serum can significantly reduce background signal and improve the signal-to-noise ratio.

The present application relates to the use of a miRNA combination in the preparation of a non-invasive diagnostic reagent for prostate cancer. Specifically, the present application provides the use of a miRNA combination as a diagnostic marker for prostate cancer, which includes multiple of hsa-miR-21-5p, hsa-miR-100-5p, hsa-miR-141-3p, hsa-miR-375-5p, and hsa-let-7f-5p. The expression levels of the miRNA combination in serum are detected by a method based on rolling circle amplification and single-strand cleavage protein, and a diagnostic model is constructed by combining a machine learning algorithm, significantly improving the diagnostic accuracy of prostate cancer. The method of the present application is a non-invasive, rapid, accurate and low-cost method for detecting prostate cancer, and has a broad application prospect.

The invention is directed to a bispecific chimeric antigen receptor, comprising: (a) at least two antigen-specific targeting regions; (b) an extracellular spacer domain; (c) a transmembrane domain; (d) at least one co-stimulatory domain; and (e) an intracellular signaling domain, wherein each antigen-specific targeting region comprises an antigen-specific single chain Fv (scFv) fragment, and binds a different antigen, and wherein the bispecific chimeric antigen receptor is co-expressed with a therapeutic control. The invention also provides methods and uses of the bispecific chimeric antigen receptors.

The application belongs to the technical field of cell engineering, and particularly relates to a bispecific antibody targeting CD22 and CD19, a chimeric antigen receptor thereof and application. In the bispecific antibody, a CD22 light chain and a CD22 heavy chain are connected through a linker 4 to form a CD22 single-chain antibody, and a CD19 light chain and a CD19 heavy chain are connected through a linker 4 to form a CD19 single-chain antibody, which comprises one of the following structures: (1) the CD22 single-chain antibody and the CD19 single-chain antibody are connected through a linker 1 or a liner 2; (2) the two ends of the CD19 single-chain antibody are connected with the CD22 light chain and the CD22 heavy chain through a linker 3, respectively. The chimeric antigen receptor provided by the application can simultaneously target two different tumor antigens, improve the killing of tumor cells, reduce the probability of immune escape, and reduce the tumor recurrence rate after CAR-T treatment.

The application provides a single-chain super-small fluorescent polymer nanodot modified Bacillus subtilis derived nanometer calcium carbonate composite material (SC-PNPs@CaCO3) as a high-sensitivity fluorescent sensing platform for choline acetyltransferase (ChAT) activity detection. The composite material is synthesized through biological mineralization of Bacillus subtilis, the free radical polymerization method is used to prepare the SC-PNPs with positive charges on the surface, and a stable fluorescent sensing system is constructed through self-assembly. Coenzyme A (CoA) is generated in the ChAT catalytic reaction, which can significantly quench the fluorescent signal of the material, realize high-sensitivity detection of the ChAT activity change, and the detection limit reaches 0.00074 mU / mL. The fluorescent sensing platform can realize real-time monitoring of the ChAT activity change, and reveal the regulation effect of the cholinergic signal pathway in the repair process in a skin repair model. The application provides an innovative technical means for choline acetyltransferase activity analysis, and provides a new research direction for exploring the function of the cholinergic system in skin repair and other physiological and pathological processes.

This invention discloses the use of an anti-PEG single-chain antibody, PEG-scFv, in conjunction with indicator liposomes for the separation and quantitative analysis of loaded and free drugs contained in small-volume biological samples containing trace amounts of PEGylated nanodrugs. The invention also discloses a method for separating and quantitatively analyzing trace amounts of loaded and free drugs in small-volume samples using PEG-scFv and indicator liposomes, comprising: adding PEG-scFv and indicator liposomes to the sample, incubation, centrifugation, aspirating the supernatant, and detecting the content of free drug in the supernatant. This separation method achieves both efficient separation of trace amounts of PEGylated nanodrugs in small-volume samples and accurate quantification of loaded and free drugs. Furthermore, the method uses the sedimentation efficiency of the indicator liposomes to indicate the separation efficiency of the PEGylated nanodrugs, meeting the consistency detection requirements for samples with unknown concentrations.

The present application belongs to the technical field of bioengineering. The present application relates to a recombinant protein, which comprises two dominant antigen epitopes of feline calicivirus (FCV) protein. In order to improve the yield of the recombinant protein in a prokaryotic expression system, the amino acid sequence of the recombinant protein is converted into a corresponding nucleotide sequence by using E. coli preferred codons, the nucleotide sequence is chemically synthesized, and a recombinant expression vector is constructed. The present application also relates to the establishment of a phage library by immunizing mice with the recombinant protein, the screening of a single-chain antibody (scfv) sequence corresponding to the feline calicivirus recombinant protein by panning, the construction of a complete mouse IgG1 antibody sequence expression vector from the obtained scfv sequence, the expression of a monoclonal antibody by instantaneously transferring HEK293F cells, the purification of the monoclonal antibody, and the labeling of europium ions (Eu 3+ ) in the monoclonal antibody, respectively. The optimal monoclonal antibody pairing combination is determined by orthogonal experiments, and the monoclonal antibody can be used for the early diagnosis of upper respiratory tract infection in cats.

The present disclosure provides a single-chain T-cell modulatory polypeptide (TMP) that includes an NKG2 peptide, an HLA-E heavy chain polypeptide, a beta-2 microglobulin polypeptide, and an immunoglobulin Fc polypeptide. The TMP can also include an immunomodulatory polypeptide and / or a cancer-targeting polypeptide. The TMPs are useful in methods of modulating an immune response, which methods are also provided.