122 results about "Phage Display Techniques" patented technology

The invention discloses a monoclonal antibody H2 specifically bound with a lung cancer marker CYFRA21-1 and application, and relates to the technical field of biological medicines. The method comprises the following steps: firstly, screening from a Tomlinson I + J phage library by utilizing a phage display technology to obtain a lung cancer marker CYFRA21-1 monoclonal antibody H2, and the antibody has a heavy chain complementary determining region and a light chain complementary determining region which are formed by novel amino acid sequences and can be specifically combined with CYFRA21-1 protein; the monoclonal antibody H2 of the lung cancer marker CYFRA21-1 has the characteristics of high affinity, high specificity, stability and the like, and the monoclonal antibody H2 of the lung cancer marker CYFRA21-1 not only can efficiently and accurately detect the concentration of CYFRA21-1 protein, but also can be used for immunostaining of lung cancer tumor cells and lung cancer tissues in pathological diagnosis, and has important significance for detecting the concentration of CYFRA21-1.

The invention relates to a method for screening a human-mouse cross MR1 restriction high-affinity TCR (T cell receptor) by using a phage display technology, and relates to the technical field of T cell receptors. According to the invention, a phage display technology is utilized to prepare an MAIT cell single-chain T cell receptor (scTCR) phage library, the scTCR phage library is subjected to biological elutriation through an MR1 / 5-OP-RU tetramer, and Phage-ELISA identifies that TCR6 clone (TCR6) has specific binding with human and mouse MR1 / 5-OP-RU and has higher affinity compared with other clone.

The invention discloses preparation and application of a biological material for targeted therapy of osteoarthritis, and the preparation method comprises the following steps: according to a phage screening principle, after phages are combined with cartilage cells, washing off phages which are not combined with the cartilage cells, are not firmly combined with the cartilage cells and are non-specifically combined with the cartilage cells; eluting the specifically bound bacteriophage with glycine acid; after the specific binding bacteriophage is amplified by escherichia coli, screening is carried out by utilizing a bacteriophage display technology and cartilage cells; the preparation method comprises the following steps: preparing a small-size porous microgel loaded drug by adopting a multiple Pickering emulsion template method; and grafting the screened cartilage targeting peptide to the surface of the porous hydrogel microsphere to obtain the HS-SP microsphere. According to the invention, the problem of OA targeting can be effectively solved by adopting a phage display technology, and a drug is precisely targeted and delivered to an OA lesion region; the porous hydrogel microspheres with small particle size are prepared by adopting a multiple Pickering emulsion method, so that the use of a large number of surfactants is avoided, and the efficient loading and release of drugs can be realized.

The invention discloses an application of an OPA1-enriched exosome in preparation of a medicine for improving mitochondrial dysfunction. The preparation method comprises the following steps: screening by adopting a phage display technology to obtain a CD9-targeted polypeptide P13, forming a fusion protein OPA1-P13 from the P13 and OPA1, constructing a recombinant plasmid for expressing the fusion protein, transfecting a cell for co-expression, and extracting the exosome, thereby obtaining the OPA1-enriched exosome OPA1-Exo. The OPA1-Exo is used for treating the mitochondrial dysfunction, and the result shows that the OPA1-Exo can activate antioxidant enzyme to reduce the ROS level in cells, promote an oxidative phosphorylation pathway to improve the ATP content of mitochondria, obviously improve a mitochondrial network and recover the mitochondrial dysfunction, so that the obvious mitochondrial function repair potential is shown. According to the invention, the OPA1 protein is successfully used for treating the mitochondrial dysfunction in the form of the exosome.

The invention relates to the technical field of biology, in particular to an anti-PM-Scl monoclonal antibody and application thereof. According to the invention, a PM-Scl recombinant protein is used as an antigen, and a phage display technology is used for screening to obtain the monoclonal antibody of the human PM-Scl. The monoclonal antibody is used as a raw material of a PM-Scl-IgG detection reagent quality control product, is good in universality and stability, can replace a clinical positive sample and be used as a quality control product of a third-party detection laboratory, saves the detection cost, is a human source, is closer to a natural sample in property compared with a human-mouse chimeric antibody, and has a good application prospect. Some matrix effect problems can be avoided.

The invention belongs to the technical field of biological medicine, and particularly relates to a nanometer antibody aiming at feline infectious peritonitis virus and application of the nanometer antibody. According to the invention, specific nano antibodies 2S-29 and 2S-39 of two novel FIPV S proteins are obtained by screening through a phage display technology, and a double-antibody sandwich ELISA method for detecting FIPV S antigens is provided. The two antibodies have excellent specificity and affinity to the feline infectious peritonitis virus FIPV S protein. A double-antibody sandwich ELISA method established based on the nano antibody has good specificity, repeatability and sensitivity, the lowest detection limit reaches 4 ng / mL, the FIPV S antigen can be rapidly and accurately detected, and a reliable means is provided for early diagnosis of feline infectious peritonitis.

The invention belongs to the technical field of immunology, and provides a recombinant C-type single-domain nano antibody combined with CDH17 and application of the recombinant C-type single-domain nano antibody. According to the invention, an antibody with binding capacity to CDH17 is screened by using a phage display technology, then m01s is used as a skeleton recombinant antibody, and the C-type single-domain antibody with high affinity activity is obtained through antibody recombination. The obtained recombinant C-type single-domain nano antibody combined with CDH17 can be used as an indicator for diagnosis and prognosis of cancers, and is expected to become a candidate drug of CAR-T drugs for gastric cancer, breast cancer, colon cancer and the like due to high affinity to tumor cells.

The invention belongs to the technical field of immunodetection, and discloses a preparation and characterization method of a rabbit-derived single-chain antibody for detecting a liver cancer marker GPC3. The rabbit-derived single-chain antibody has the characteristics of small volume, high specificity, high stability and good sensitivity, and can be used in an immunoassay method for detecting a liver cancer marker GPC3; according to the method, a specific rabbit single-chain antibody is obtained through screening, expression and purification by using a phage display technology, a GPC3 antigen is coated on a solid-phase carrier, the rabbit single-chain antibody is used as a detection carrier, the binding force of the single-chain antibody and the antigen is identified, and the method is high in specificity, good in sensitivity and high in specificity. A novel single-chain antibody with high specificity and high sensitivity is provided for detection of GPC3 in serum.

The invention belongs to the technical field of biological medicines, and particularly relates to an anti-B7H3 nano antibody and application thereof. The invention provides an anti-B7H3 nano antibody and application thereof, aiming at the problems that the existing B7H3 antibody is mainly a mouse monoclonal antibody and is large in molecular weight, poor in tissue penetrability, not easy to engineer and the like. According to the invention, a human B7H3 target spot is taken as an antigen, 15 anti-B7H3 specific nano antibodies are screened through a phage display technology, and the nano antibodies can be specifically combined with the B7H3 antigen, are high in specificity and affinity, and also have the advantages of small molecular weight, high stability, easiness in transformation, high solubility and the like. Meanwhile, the anti-B7H3 nano antibody and the anti-CD3 antibody OKT3scFv are connected in series to prepare the B7H3 / CD3 bispecific antibody, in-vivo and in-vitro experiments show that the B7H3 / CD3 bispecific antibody has a good anti-tumor effect, and the B7H3 / CD3 bispecific antibody can be applied to preparation of anti-tumor drugs and has a wide anti-tumor effect.

This invention belongs to the field of microcystin-LR detection technology, providing anti-microcystin-LR nanobodies and their applications. The anti-microcystin-LR nanobodies have amino acid sequences as shown in SEQ ID NO.1, SEQ ID NO.2, or SEQ ID NO.3. This invention uses phage display technology to screen for nanobodies with high affinity and high specificity for recognizing microcystin-LR, exhibiting advantages such as strong stability and ease of large-scale expression. Various detection tools can be designed based on anti-microcystin-LR, including test strips, immunoassay kits, immunoaffinity columns, biosensor detection chips, or detection probes. This provides novel antibody resources for rapid monitoring of microcystin-LR and lays a material foundation for developing low-cost, high-performance immunoassay technologies, which is of great significance for promoting the monitoring and control of microcystin-LR pollution.

The present invention discloses a monoclonal nano antibody for recognizing Epstein-Barr virus gp42 protein, its application and monoclonal nano antibody library, and relates to the technical field of phage display of nano antibodies. The monoclonal nano antibodies EBV-gp42-1023, EBV-gp42-1024 or EBV-gp42-1025 provided by the present invention, and related biomaterials, pharmaceutical compositions or detection products, have very high affinity and specificity for Epstein-Barr virus gp42 protein, and can be widely used in detecting the expression of Epstein-Barr virus gp42 protein, or detecting the occurrence and development process of Epstein-Barr virus-induced tumors, or evaluating the efficacy of drugs for treating Epstein-Barr virus-induced tumors, or treating Epstein-Barr virus-induced tumors.

The invention discloses a human-derived hysteromyoma targeting peptide and application thereof, the sequence of the targeting peptide is selected from SEQ ID NO: 1 to SEQ ID NO: 5, and the SEQ ID NO: 1 (VN12) is the most preferable. The fusion protein is obtained by screening through a phage display technology, and has high hysteromyoma targeting specificity and low immunogenicity. The targeting peptide can be coupled with nuclide or fluorescent dye, and is used for accurate PET imaging, fluorescent diagnosis and intraoperative navigation of hysteromyoma. Experiments prove that the molecular marker is specifically enriched in tumor tissues (the tumor / muscle ratio is 60), the biological safety is good, the problems of insufficient diagnosis sensitivity and large treatment wound in the prior art are solved, and the molecular marker has a great application prospect in diagnosis and treatment of hysteromyoma.

The invention belongs to the technical field of biological medicine, and particularly relates to an anti-human DCBLD2 nano antibody and application thereof. The anti-DCBLD2 nano antibody is successfully screened by using a phage display technology, and the nano antibody can be specifically combined with human DCBLD2 protein, can be prepared into an immune complex or a pharmaceutical composition for preventing or treating DCBLD2 target related diseases, and has a wide application prospect.

The application belongs to the technical field of biological medicine, and discloses a cyclic peptide molecule specifically targeting CD5 and purposes thereof. The surface receptor CD5 of immune cells (T cells, B cells) has been proved to be a potential tumor treatment target, the present application uses the phage display technology widely applied to the development of polypeptide or antibody drugs to carry out multiple rounds of in vitro screening on human CD5 protein, and obtains a cyclic peptide molecule with drug potential and capable of specifically combining with CD5 through second-generation sequencing. The cyclic peptide molecule contains two cysteines, is modified into a ring through a specific chemical crosslinking agent, and the affinity of the cyclic peptide molecule to CD5 is determined to be in the low micromolar level through surface plasmon resonance technology, thereby laying a foundation for the development of a CD5-targeting inhibitor.

This invention belongs to the field of biomedical technology, specifically relating to a colorectal cancer antibody and its applications. The antibody is composed of a camel-derived VHH-3 single-domain antibody and a membrane-penetrating peptide TAT fused together via a flexible linker peptide (GGGGS)3, and its amino acid sequence is shown in SEQ ID NO:9. This invention uses phage display technology to screen for high-affinity VHH-3 (KD=0.9524 nM) and further constructs the membrane-penetrating nanobody TAT-VHH-3, achieving soluble and efficient preparation using a prokaryotic expression system. Experiments confirm that TAT-VHH-3 can significantly inhibit the proliferation (CCK-8 assay) and migration (scratch assay) of colorectal cancer Caco-2 cells in a concentration-dependent manner. The nanobody of this invention has advantages such as small molecular weight (approximately 16.1 kDa), strong penetrability, and high stability, providing a novel tool for tumor therapy targeting CaMKⅡα.

The present invention discloses an intestinal microbiota Bacteroides fragilis toxin-neutralizing nanobody and its application. By deeply studying the enterotoxigenic Bacteroides fragilis and the toxin BFT secreted by it in the bodies of patients with colorectal cancer and breast cancer, nanobodies against BFT are expressed by phage display technology; nanobodies with high binding affinity to the antigen are screened out through biopanning technology; a nanobody with the ability to neutralize the toxicity of BFT, also known as Nb2.43, is obtained. The present invention expresses and optimizes to obtain a BFT nanobody protein with high affinity, stability and uniformity, opening up a new field for the treatment of colorectal cancer and breast cancer with nanobodies targeting BFT, and having good research value and application prospects.

The application relates to the field of antibody technology, in particular to an ACTH recombinant rabbit monoclonal antibody, a preparation method and application thereof. The ACTH recombinant rabbit monoclonal antibody is screened by using phage display technology. The monoclonal antibody has high specificity, high detection sensitivity and good affinity, and solves the problems of low content of adrenocorticotropic hormone in samples and great detection difficulty. Through identification, the monoclonal antibody has good stability and high accuracy when repeatedly detected.

The present invention discloses an EBV-related gastric cancer diagnostic marker and its application, wherein the diagnostic marker is selected from at least one of LF2 protein and BFRF3 protein. The present invention is the first to screen and obtain two diagnostic markers for EBV-related gastric cancer (EBVaGC) based on proteome chips and phage display technology - LF2 protein and BFRF3 protein, which can efficiently and accurately diagnose EBVaGC. In particular, the AUC of the combined diagnosis of these two proteins can reach 0.911, which is significantly better than the AUC of the individual diagnoses. And when diagnosing EBVaGC based on these two proteins, no tissue sample is required, only a serum sample is required, and it can be detected during a routine blood test. Therefore, compared with existing diagnostic methods, the present invention has the characteristics of high accuracy, small limitations, and low cost, reduces the cost of medical treatment for patients, is conducive to the realization of universal screening of the population, simplifies the diagnosis process of EBVaGC, and significantly improves the treatment efficiency.

A heavy-chain variable region sequence of the nanobody CPV-VHH-H1 has the amino acid sequence set forth in SEQ ID NO: 1; and a gene encoding the nanobody CPV-VHH-H1 has the nucleotide sequence set forth in SEQ ID NO: 2. A nanobody immune library of the CPV is constructed by a phage display technology, a specific anti-CPV nanobody CPV-VHH-H1 is obtained by screening, and it is verified by experiments that the nanobody may specifically bind to the CPV. A new nanobody preparation for use in clinical diagnosis and treatment of the CPV can be developed, and a certain theoretical reserve is provided for applying the nanobody to the field of veterinary biological products.

The application relates to the technical field of biological medicine, and provides an anti-GPC3 full humanization single-domain antibody and application thereof. The application uses phage display technology to obtain one anti-human GPC3 specificity full humanization monoclonal single-domain antibody through antibody screening by targeting human GPC3 protein. The obtained single-domain antibody can be specifically combined with human GPC3, and can effectively inhibit the nuclear translocation of YAP, provides an effective alternative antibody drug for GPC3-targeted HCC treatment, and has a potential clinical application prospect.

The application belongs to the field of biological medicine, and provides an antibody combined with a specific antigen peptide of common pemphigus and application thereof, and the application uses the principle of phage display technology to perform in-vitro affinity screening on a constructed library, combines phages with target antigen Dsg3, elutes non-specific binders, retains phages combined with the antigen closely, and then amplifies the phages and performs multiple rounds of affinity enrichment; multiple rounds of panning are performed, and after 3-5 rounds of screening, phage clones with strong Dsg3 binding capacity and high specificity can be obtained.

The invention belongs to the technical field of biological medicines, and particularly relates to an anti-CD22 nano antibody as well as a preparation method and application thereof. The anti-CD22 nano antibody comprises a framework region and a complementarity determining region, wherein the complementarity determining region comprises CDR1 (SEQ ID NO: 1), CDR2 (SEQ ID NO: 2) and CDR3 (SEQ ID NO: 3). The preparation method comprises the following steps: immunizing alpaca by using Human CD22 / His protein, extracting RNA (Ribonucleic Acid) from PBMC (Peripheral Blood Mononuclear Cell) and reversely transcribing into cDNA (Complementary Deoxyribose Nucleic Acid), constructing a phage display library, then performing CD22 antigen protein elutriation to obtain positive clone, and after sequencing analysis, expressing the antibody by using a mammalian cell expression system. The optimized phage display technology is adopted, the screening period is short, and the obtained nano antibody is small in molecular weight, stable in structure and high in affinity to CD22. The antibody can be efficiently expressed through mammalian cells, and natural modification activity is reserved.

This invention provides an anti-trimethoprim single-chain antibody for detecting trimethoprim-like drug residues, its preparation method, and its application. The amino acid sequences of the single-chain antibody are shown in SEQ ID3, SEQ ID4, and SEQ ID6. Compared with traditional monoclonal antibodies, the single-chain antibody has lower production costs, is simpler to prepare, and is easier to manipulate at the gene level. This invention utilizes trimethoprim-specific monoclonal hybridoma cells and employs phage display technology to screen for high-affinity single-chain antibodies. A single-chain antibody mutant with even higher affinity was successfully obtained through computer simulation combined with site-directed mutagenesis. Based on this mutant, an ic-ELISA was established for the detection of trimethoprim residues in animal-derived foods, meeting the needs of practical applications.

The present application relates to the field of biotechnology, in particular to a kind of hyaluronidase binding cyclic peptide, the cyclic peptide is obtained by screening phage display technology, its structure includes X 1(n) CX3X4X5X6X7X8X9X 10 CX 12(m) It has the efficacy of inhibiting hyaluronidase activity, higher transdermal absorption capacity and / or anti-inflammatory, can be used as hyaluronidase (especially PH20) inhibitor, applied in the fields of medicine, food and cosmetics.

The application discloses a single-chain antibody, the amino acid sequence of which is shown as SEQ ID NO:1, and the nucleotide sequence of which is shown as SEQ ID NO:2. The single-chain antibody against rabies virus is screened from an antibody library of immunized mice by using a phage display technology. Compared with a traditional cell fusion technology, the phage display technology is simple in operation, low in cost, short in time, and high in antibody discovery and screening efficiency. The single-chain antibody obtained by the application is low in molecular weight and immunogenicity, high in antibody specificity and affinity, and has the activity of neutralizing rabies virus, thereby providing a new idea for basic research of rabies virus and development of therapeutic drugs.

Neuroendocrine tumors (NETs) are highly heterogeneous tumors derived from neuroendocrine cells. The synaptic vesicle 2 (SV2) protein is a novel marker of NETs and has three subtypes of SV2A, SV2B and SV2C, the SV2C structure is more stable than the other two subtypes, and the maximum cavity ring L4 is the most important binding region with toxins. The nano antibody of the extracellular soluble part L4 of the SV2 is obtained by screening through a phage display technology, the obtained nano antibody is labeled by fluorescein, chemical toxin small molecules or nuclide to prepare a probe molecule, and the result can specifically bind to neuroendocrine tumor cells. The invention provides a nano antibody of an extracellular soluble part L4 of SV2. The nano antibody can be applied to preparation of a preparation for diagnosing and treating neuroendocrine tumors.

The invention belongs to the field of biological medicine, and provides an antibody combined with pemphigus vulgaris specific antigen peptide and application of the antibody. According to the principle of a phage display technology, a constructed library is subjected to in-vitro affinity screening, phages are combined with a target antigen Dsg3, non-specific binders are eluted, the phages tightly combined with the antigen are reserved, and the pemphigus vulgaris specific antigen peptide is obtained. Then, amplifying and carrying out affinity enrichment for multiple rounds; and carrying out multiple rounds of panning, and after 3-5 rounds of screening, obtaining phage clones with strong binding capacity with Dsg3 and high specificity.

The application belongs to the technical field of bioengineering, and relates to a fluorescent probe for detecting neuron-specific enolase and application. The application screens a specific binding short peptide E for neuron-specific enolase by using phage display technology, inserts a circular fluorescent protein R-GECO1 into the short peptide E, divides the E peptide chain into two parts E1 and E2, and forms a probe structure in the form of E1-R-GECO1-E2. The fluorescent probe protein provided by the application has relatively small molecular weight and is easy to mature, has large fluorescence dynamic change, has good specificity, and can detect neuron-specific enolase in and outside cells.

The invention relates to the technical field of antibodies, in particular to a humanized anti-beta2-GP1 monoclonal antibody as well as a preparation method and application thereof. The full-length IgG / IgM / IgA type antibody sequence is obtained by constructing a natural full human source library, adopting a phage display technology, screening out a high-affinity antigen binding fragment, and assembling a variable region sequence and an IgG / IgM / IgA constant region. An anti-beta2-GPI antibody is expressed through transient transfection, a protein A / L affinity purification method is adopted for preparation, and binding activity determination is carried out. Results show that the specific beta2-GPI antibody obtained by the method is high in antigen binding activity, high in specificity and good in affinity.

The invention discloses a CD147 nano antibody G45 as well as a preparation method and application thereof, and belongs to the technical field of molecular biology and immunology. A phage display technology is used for screening from a natural alpaca nano antibody phage library, finally three nano antibody genes are selected and cloned into pCold II, and prokaryotic expression recombinant plasmids are successfully constructed; the method comprises the following steps: carrying out inducible expression for 20 hours under the conditions that the final concentration of IPTG (isopropyl-beta-d-thiogalactoside) is 0.2 mmoL / L, the temperature is 16 DEG C and the speed is 100 rpm / min, carrying out nickel column purification, and then carrying out Coomassie brilliant blue dyeing and Western blot identification, so as to determine and obtain the CD147 nano antibody which is named as A8, B8 and G45; the binding activity of the three nano antibodies and the CD147 protein is detected through indirect ELISA (enzyme-linked immuno sorbent assay). The result shows that the CD147 antigen and the three nano antibodies have binding activity and are dose-dependent. Under the same conditions, when the incubation concentration of the nano-antibody is as low as about 0.625 g / mL, the G45 binding effect is good, the amino acid sequence of the nano-antibody is shown as SEQ ID NO.7, and the nucleotide sequence of the nano-antibody is shown as SEQ ID NO.10. The invention provides a more accurate method for high-sensitivity detection of CD147.