127 results about "Phage display" patented technology

The invention belongs to the technical field of food and feed analysis and detection, and particularly provides a nano antibody for detecting bacitracin methylene salicylate, and the nano antibody comprises a nano antibody BMD-BA19 and a nano antibody BMD-CA45; the amino acid sequence of a VHH chain of the nano antibody BMD-BA19 is as shown in SEQ ID NO: 1, and the amino acid sequence of a VHH chain of the nano antibody BMD-CA45 is as shown in SEQ ID NO: 2. The bacitracin methylene salicylate serves as a research object, after the bacitracin methylene salicylate is coupled with protein, an alpaca is immunized, two nano antibodies resisting bacitracin methylene salicylate are obtained through screening with a phage display technology, and a material is provided for establishing a bacitracin methylene salicylate detection method.

The invention discloses a nano antibody targeting GPRC5D as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The nano antibody is generated by mixed immunization of a GPRC5D polypeptide antigen with a sequence shown as SEQ ID NO: 1 and a GPRC5D protein antigen with a sequence shown as SEQ ID NO: 2. The preparation method comprises the following steps: mixing the GPRC5D polypeptide antigen and the GPRC5D protein antigen to immunize alpaca, sampling blood, constructing an antibody library, screening a phage display antibody library, expressing protein, and selecting the nano antibody with the highest affinity. The invention further provides a radionuclide labeled molecular probe, the nano antibody is small in molecular weight and can recognize a large number of epitopes, and the molecular probe prepared from the nano antibody can obtain high-quality images, is used for preparing products for developing multiple myeloma or detecting GPRC5D expression level and has wide application prospects.

The present invention relates to the technical field of molecular biology, and in particular, to an anti-CD40 nanobody, and a preparation method therefor and use thereof. The anti-CD40 nanobody provided by the present invention has three unique complementarity determining regions, namely CDR1, CDR2, and CDR3. The present invention further provides a sequence encoding the nanobody or its VHH chain, a corresponding expression vector, and a host cell capable of expressing the nanobody. The present invention includes immunizing an alpaca by constructing a CD40 antigen to obtain a PBMC of the alpaca and constructing a vector; screening an anti-CD40 antibody through phage display; and finally obtaining a highly sensitive and specifically binding antibody through functional assays and sequencing result analysis. The nanobody provided by the present invention can specifically recognize and bind to CD40.

The present invention relates to the field of biotechnology. Specifically, the present invention provides a monoclonal antibody, preparation method, and application of tyrosine iodinated EGFR protein. The present invention prepares tyrosine iodinated EGFR protein antigen peptides, immunizes mice to prepare monoclonal antibodies, and utilizes phage display technology to select and express relevant antibodies using expression vectors. ELISA screening and immunohistochemistry and cell fluorescence detection yield two highly sensitive and specific monoclonal antibodies, M018 and M025. The monoclonal antibodies obtained by screening can be used for the detection and monitoring of tyrosine iodinated modification at position 845 of the EGFR protein, providing support for tumor diagnosis and the development of anti-tumor drugs.

The invention discloses a preparation method and application of a cat main allergen FEL-1 nano antibody FEL-1-Nb, and relates to the technical field of antibody preparation, and the preparation method comprises the following steps: preparing an immunogen; animal immunization and library construction; constructing and panning a phage display library; screening and identifying positive clone; and expression and purification of the nano antibody: cloning the positively cloned VHH gene to an expression vector such as a pET series, transforming an expression strain such as escherichia coli BL21 (DE3), carrying out IPTG induced expression, and purifying through an inclusion body purification method to obtain a high-purity nano antibody protein. The nano antibody FEL-1-Nb provided by the invention can be specifically bound with a cat main allergen FEL-1 protein, the unique amino acid sequence of the nano antibody FEL-1-Nb is shown as SEQIDNO: 1, high-affinity binding with the FEL-1 protein is ensured, the positive clone binding rate is high through phage display panning and ELISA verification, the phage recovery rate after the third round of panning is increased by more than 1000 times compared with the first round of panning, and the positive clone binding rate is higher than that of the first round of panning. Therefore, the nano antibody has excellent targeted recognition capability.

This invention discloses a polypeptide that specifically binds to the coronavirus S protein, its encoding gene, and its applications, belonging to the field of molecular biology. The polypeptide specifically binding to the coronavirus S protein provided by this invention contains the following amino acid sequence: HFVKTPARWAWG, obtained through screening using phage display technology. The polypeptide provided by this invention can specifically bind to the coronavirus S protein and specifically block the binding of the coronavirus S protein to ACE2, thereby inhibiting coronavirus infection and can be used to prepare anti-coronavirus drugs.

This invention provides a peanut allergen peptide, its screening method, and its application. Phage display technology is used to screen for conformational epitopes of Ara h5 and Ara h8 proteins. Polyclonal IgE and IgG antibodies against the target proteins are prepared by immunizing BALB / c mice with Ara h5 and Ara h8 proteins as antigens. ELISA and dotted immunoblotting detection revealed that the titers of polyclonal IgG and IgE against Ara h5 and Ara h8 proteins were 1:64000 and 1:320, respectively, and the antibodies exhibited specificity. Using the antibodies as targets, phage display technology screened the conformational epitope sequences of Ara h5 protein as WETIYSR and FHWWYLK. The conformational epitopes of Ara h8 protein were FPYMKFV, FPYMKFR, SMFARID, and SFHWWLF.

The invention relates to the technical field of biology, and aims to provide an M13 recombinant bacteriophage with PRRSV broad-spectrum neutralizing activity for displaying NPC and application of the M13 recombinant bacteriophage. A nano antibody-CD163 peptide fragment conjugate NPC shown as SEQ ID NO: 1 is displayed on the surface of the N end of capsid protein III of the M13 recombinant phage, 1-3 NPC molecules are contained on the surface of each phage particle, and the phage is used for neutralizing PRRSV (Porcine Reproductive and Respiratory Syndrome Virus). The recombinant bacteriophage provided by the invention is easy to prepare, extremely high in virus titer and excellent in thermal and pH stability; in Marc-145 and PAMs cells, the compound shows strong neutralizing activity on PRRSV II type 1, 3, 5 and 8 pedigree, so that the compound has broad-spectrum neutralizing capacity; the compound can be used as a PRRSV prevention and control preparation, and a new thought is provided for development of safe and efficient novel antiviral drugs or biological preparations.

The invention belongs to the technical field of tumor immunotherapy, and relates to an anti-Claudin18.2 single-domain antibody, a multifunctional fusion protein, a recombinant natural killer cell (CT-CAR-NK), and preparation and application thereof. Through alpaca immunization and phage display library construction and panning, the single-domain antibody VHH-C18.2-1 specifically combined with Claudin18.2 is obtained, and the amino acid sequence of the single-domain antibody VHH-C18.2-1 is SEQ ID NO: 1. The amino acid sequence of the designed fusion protein is SEQ ID NO: 3, the fusion protein sequentially comprises a VHH-C18.2-1, a flexible Linker, a TGF-beta RII extracellular domain, a CD8alpha hinge region, a CD8alpha transmembrane region, a 4-1BB intracellular domain and a CD3zeta intracellular domain from the N end to the C end, and the fusion protein has the functions of targeting, resisting TGF-beta inhibition and activating signals. The fusion protein gene transfects human peripheral blood CD56 + CD3-NK cells through lentivirus to obtain CT-CAR-NK, in-vitro verification shows that the CT-CAR-NK still keeps efficient killing in an immunosuppression environment, tumor growth can be remarkably inhibited in vivo, the lifetime can be prolonged, and a safe and efficient scheme is provided for Claudin18.2 positive solid tumor treatment.

The invention belongs to the technical field of in-vitro diagnosis, and particularly relates to an anti-mycoplasma pneumoniae antibody, a mycoplasma pneumoniae detection quality control product and a preparation method and application of the mycoplasma pneumoniae antibody and the mycoplasma pneumoniae detection quality control product. The invention provides an anti-mycoplasma pneumoniae antibody which comprises a light chain variable region and a heavy chain variable region, and the light chain variable region and the heavy chain variable region are specifically combined with a C-terminal immunodominance region of mycoplasma pneumoniae P1 protein. Wherein the C-terminal repeat region of the mycoplasma pneumoniae P1 protein is a core immunogenicity region, has high conservative property and strong immunogenicity, can induce a host to generate a specific IgM antibody in the early stage of infection, and is an ideal antibody target. The anti-mycoplasma pneumoniae antibody is obtained by screening through a phage display technology, has high affinity and high specificity, can effectively recognize the C-terminal immunodominance region of the mycoplasma pneumoniae P1 protein, and provides a reliable detection tool for early diagnosis of mycoplasma pneumoniae.

The invention relates to the technical field of medical supplies, and discloses an orally disintegrating film for promoting remineralization of enamel and a preparation method thereof.The orally disintegrating film takes hydroxypropyl methyl cellulose, microcrystalline cellulose, starch and the like as film base materials and has the advantages of being thin, light, good in flexibility, free of toxicity and stimulation, good in biocompatibility and biodegradability and the like; and the composition can be quickly dissolved in the oral cavity to take effect. The device has the advantages of quick effect, convenience in carrying, easiness in use and the like. The orally disintegrating film serves as a direct carrier of an inducer for inducing remineralization of enamel, so that the orally disintegrating film becomes a more effective transmission system. A bacteriophage display technology is utilized, a peptide fragment with strong adsorption capacity with HAP is integrated into a carrier bacteriophage DNA, and purification is performed after large-scale amplification, so that the peptide fragment is used as an important effective component of the orally disintegrating film.

The invention belongs to the technical field of biological pharmacy, and particularly discloses a Siglec-15 affinity peptide and application thereof. According to the invention, a phage display heptapeptide library high-throughput screening technology is adopted, Siglec-15 affinity peptide 7-8 is obtained through multiple rounds of screening, and the Siglec-15 affinity peptide 7-8 is optimized and modified to obtain a reversed sequence flip peptide d-7-8. Experimental results show that the Siglec-15 affinity peptide can be used for affinity of Siglec-15 protein and blocking the interaction between Siglec-15 and a ligand Sialyl-Tn of Siglec-15, so that the anti-tumor effect is achieved; an in-vitro cell level experiment shows that the Siglec-15 affinity peptide has a relieving effect on inhibition of Siglec-15 on CD8 + T cell proliferation, has an obvious immune activation effect, does not have obvious toxic or side effects, can be used for preparing an anti-tumor drug or a detection reagent taking Siglec-15 as a target spot, and has a relatively good medical application prospect.

The present application relates to a kind of artificially synthesized recombinant monoclonal antibody with human antibody amino acid sequence as framework.The present application uses phage display technology, and obtains one monoclonal antibody H11 from Tomlinson I+J phage library, and these new amino acid sequences constitute CDR1, CDR2 and CDR3 region, can be combined with A29L protein specific binding.It shows good stability, high specificity and high affinity and other characteristics, provide tool support for subsequent monkeypox virus diagnosis and detection with A29L protein as target.

The invention discloses a cat parvovirus VP2 protein nano antibody as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The amino acid sequence of the cat parvovirus VP2 protein nano antibody is as shown in SEQ ID NO. 1 or SEQ ID NO. 2. According to the invention, an alpaca is immunized by using FPV VP2 protein, a phage display nano antibody library is constructed, and two nano antibodies 3D6 and 1B9 with strong specificity and high affinity are obtained through multiple rounds of panning and identification. Through identification, the nano antibodies 3D6 and 1B9 can be specifically combined with FPV and VP2 proteins, and have high affinity and remarkable virus neutralizing activity. The nano antibody can be used for preparing a detection reagent or kit for feline parvovirus infection, and rapid and accurate diagnosis is realized; the compound can also be used as an active component to prepare a therapeutic drug, and provides a special therapeutic scheme for FPV infection. The nano antibody provides an important material basis for developing a novel diagnostic tool and a specific therapeutic drug aiming at the feline parvovirus.

This invention discloses nanobodies against rabies virus G protein and their applications, belonging to the field of nanobody technology. The nanobodies are antibody 11G6, antibody 9F7, or antibody 9A3; the amino acid sequence of antibody 11G6 is shown in SEQ ID NO. 8; the amino acid sequence of antibody 9F7 is shown in SEQ ID NO. 13; and the amino acid sequence of antibody 9A3 is shown in SEQ ID NO. 18. This invention uses phage display technology to screen for nanobodies with high affinity and high neutralizing activity. Based on this, this invention utilizes the advantage of the small size of nanobodies and verifies their unique ability to cross the blood-brain barrier (BBB) ​​through Transwell experiments, overcoming the limitation that large molecules such as antibodies and drugs cannot cross the BBB to enter the brain. This invention provides a new drug option for the treatment of rabies virus and has significant clinical implications.

The application belongs to the technical field of biology and particularly relates to a nano antibody against African swine fever virus K205R protein and application. The application provides a nano antibody KR-6 against African swine fever virus K205R protein, wherein the nano antibody KR-6 is obtained through construction and screening of a phage display nano antibody library, is a heavy chain antibody variable region fragment, and has an amino acid sequence shown in SEQ ID NO. 1. The nano antibody has high affinity to African swine fever virus K205R protein and African swine fever virus, can specifically recognize African swine fever virus, and can be used for diagnosis, prevention or treatment of African swine fever.

The invention discloses a screening method and application of a shark-derived anti-idiotypic nano antibody, and belongs to the technical field of biological pharmacy and vaccines. The screening method comprises the following steps: immunizing adult shark by using a target pathogen specific antigen, collecting peripheral blood, separating lymphocytes, extracting RNA (Ribonucleic Acid), carrying out reverse transcription to synthesize cDNA (Complementary Deoxyribonucleic Acid), amplifying a nano antibody variable region fragment of a shark IgNAR (Immunoglobulin Nucleic Acid Receptor) gene, and constructing a phage display gene bank of which the storage capacity is greater than or equal to 1 * 10 < 8 > CFU and the recombination rate is greater than or equal to 85%; a variable region of a target pathogen specific antibody is taken as a target, specific bacteriophage is enriched through 3-4 rounds of gradient washing and elutriation (Tween-20 concentration and washing time are gradually improved), and the shark source anti-idiotypic nano antibody with high affinity and high purity (greater than or equal to 95%) is obtained by combining ELISA screening positive clone, gene sequencing de-weighting and SPR detection (combining dissociation constant KDlt, 1 * 10 <-7 > mol / L).

The present invention discloses a monoclonal antibody H2 that specifically binds to the lung cancer marker CYFRA21-1 and its application, relating to the field of biomedicine technology. First, using phage display technology, a lung cancer marker CYFRA21-1 monoclonal antibody H2 is obtained by panning from the Tomlinson I+J phage library. The antibody has a heavy chain complementary determining region and a light chain complementary determining region composed of novel amino acid sequences, and can specifically bind to the CYFRA21-1 protein. The binding has the characteristics of high affinity, high specificity and stability. The monoclonal antibody H2 for the lung cancer marker CYFRA21-1 of the present invention can not only 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, which is of great significance for detecting the concentration of CYFRA21-1.

The present invention relates to the technical field of molecular biology, and in particular, to an anti-CD40 nanobody, and a preparation method therefor and use thereof. The anti-CD40 nanobody provided by the present invention has three unique complementarity determining regions, namely CDR1, CDR2, and CDR3. The present invention further provides a sequence encoding the nanobody or its VHH chain, a corresponding expression vector, and a host cell capable of expressing the nanobody. The present invention includes immunizing an alpaca by constructing a CD40 antigen to obtain a PBMC of the alpaca and constructing a vector; screening an anti-CD40 antibody through phage display; and finally obtaining a highly sensitive and specifically binding antibody through functional assays and sequencing result analysis. The nanobody provided by the present invention can specifically recognize and bind to CD40.

The present application belongs to the technical field of nanobody, and particularly relates to an anti-CHI3L1 nanobody, a construction method and application of a phage display library thereof. The present application discloses an anti-CHI3L1 nanobody, which is a nanobody aiming at an epitope of a CHI3L1 protein antigen, and has a VHH chain with an amino acid sequence shown as SEQ ID NO:1. The present application takes CHI3L1 as a research object, immunizes a llama after expression and purification of CHI3L1, and screens an anti-CHI3L1 nanobody by using phage display technology. The nanobody has the ability of high sensitivity and specificity in binding CHI3L1. The anti-CHI3L1 nanobody of the present application provides materials for establishing a diagnostic method and product of CHI3L1, and also provides a research direction for developing a drug for treating a disease related to CHI3L1.

The invention relates to a fusion polypeptide containing a cyclic peptide with a novel configuration and a single domain antibody, a phage display library for displaying the fusion polypeptide, a preparation method of the phage display library, and application of the fusion polypeptide and the phage display library. The phage display library has the advantages that the display efficiency of the polypeptide is improved, meanwhile, the polypeptide can be prepared by using eukaryotic cell strain expression, and the purification of a polypeptide sample is realized through a single-domain antibody, so that the polypeptide with higher purity and lower immunogenicity is screened and obtained with low cost and low pollution.

The invention relates to the technical field of biology, in particular to an anti-PLA2R fully human IgG monoclonal antibody and application thereof. The invention provides a human-derived anti-alkali phospholipase A2 receptor antibody PLA2R PL82 screened by using a phage surface display technology, the subtype of the human-derived anti-alkali phospholipase A2 receptor antibody PLA2R PL82 is IgG4, and amino acid sequences of light chain and heavy chain variable regions of the human-derived anti-alkali phospholipase A2 receptor antibody PLA2R PL82 are respectively shown as SEQ ID NO: 6 and SEQ ID NO: 7. The antibody can specifically recognize an alkali phospholipase A2 receptor, the reactivity, the stability and the matrix effect all meet the performance use requirements of a kit, and the antibody plays an important role in alkali phospholipase A2 IgG project application as a quality control product.

The invention discloses a CD147 nano antibody D2 as well as a preparation method and application thereof, and belongs to the technical field of molecular biology and immunology. The preparation method comprises the following steps: screening from an alpaca nano antibody phage library by utilizing a phage display technology, and sequencing to obtain 9 CD147 nano antibodies; 3 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 to determine and obtain the CD147 nano antibody which is named as C36, D2 and H4; 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 1.25 micrograms / mL, the D2 binding effect is the best, the amino acid sequence of the nano-antibody is shown as SEQ ID NO.8, and the nucleotide sequence of the nano-antibody is shown as SEQ ID NO.10. The invention provides a more accurate method for detecting the CD147.

The invention discloses an anti-rabies virus G protein nano antibody and application thereof, and belongs to the technical field of nano antibodies. The nano antibody is an antibody 11G6, an antibody 9F7 or an antibody 9A3; the amino acid sequence of the antibody 11G6 is as shown in SEQ ID NO. 8; the amino acid sequence of the antibody 9F7 is as shown in SEQ ID NO. 13; the amino acid sequence of the antibody 9A3 is as shown in SEQ ID NO. 18. According to the invention, the nano antibody with high affinity and high neutralizing activity is screened by a phage display technology. On the basis, the advantage that the size of the nano antibody is small is utilized, the unique ability of the nano antibody to penetrate through the BBB is verified through a Transwell experiment, and the defect that macromolecules such as antibodies and drugs cannot penetrate through the BBB to enter the brain is overcome. The invention provides a new drug choice for the treatment of rabies virus, and has important clinical significance.

The present application relates to the technical field of antibody, in particular to a recombinant dopamine rabbit monoclonal antibody and application. The present application utilizes phage display technology and alanine scanning mutagenesis technology, and screens to obtain a dopamine monoclonal antibody, compared with other antibodies and the antibody without alanine scanning mutagenesis, the affinity and specificity are strong, the detection sensitivity is high, and the consistency with the antigen detection concentration result in liquid quality is high. The antibody is used for the detection of dopamine in serum, and the test result shows that the detection sensitivity is high, the specificity is good, the repeatability is high, the stability is good, the precision is high, and the antibody has an important role for the clinical diagnosis of dopamine.

The invention discloses a nano antibody targeting human cardiac troponin I and application of the nano antibody. According to the invention, through construction of a nano-antibody phage library and a phage screening technology, four nano-antibodies targeting human cardiac troponin I and having a good pairing effect are obtained. Compared with the traditional monoclonal antibody, the bacteriophage display nano antibody has the advantages of small size, high stability, simplicity and convenience in preparation, low cost, large-scale production and the like; the probe can be used as a core detection element and is applied to the field of diagnosis of clinical rapid detection of human cardiac troponin I.

The invention discloses a rapid detection method for fusarium moniliforme based on nano-antibody targeted recognition, and relates to the technical field of plant disease detection.The detection method comprises the steps of nano-antibody screening and compound preparation, specifically, antigen protein is extracted from fusarium moniliforme, a camel is selected as an immune animal for immune treatment, and a compound is obtained; separating lymphocytes to construct a nano antibody gene pool; through nano antibody screening and compound preparation technologies, the sensitivity and the specificity of detection of the rice fusarium moniliforme are remarkably improved, antigen protein is extracted from the rice fusarium moniliforme, a camel is selected as an immune animal for immune treatment, a nano antibody gene pool is constructed, and the sensitivity and the specificity of detection of the rice fusarium moniliforme are improved. Nano antibodies with high affinity and specificity are screened out by utilizing a phage display technology, and the nano antibodies can accurately recognize and combine specific epitopes of pathogenic bacteria, so that the problem of cross reaction possibly existing in traditional antibodies is avoided.

The present invention belongs to the field of biopharmaceutical technology, and specifically discloses a TIM-3 affinity peptide and its application. The present invention obtains TIM-3 protein affinity peptide TBS-22 (as shown in SEQ ID NO.1) by screening through phage display seven peptide library high-throughput screening technology, and obtains mutant peptide TBSM-3 (isoleucine at position 6 is mutated to histidine, as shown in SEQ ID NO.2) through later optimization and transformation. The present invention shows through in vitro cell level experiments and mouse tumor-bearing experiments that TIM-3 affinity peptide can affinity TIM-3 protein, block the interaction between TIM-3 and its ligand Galectin-9, thereby exerting anti-tumor efficacy, can be used to prepare anti-tumor drugs, has good medical application prospects, and provides a new option for tumor immunotherapy.

Single-domain monoclonal antibodies that specifically bind B7H3 (also known as CD276) are described. The single-domain antibodies include camel VHH and rabbit VH domain nanobodies selected from phage display libraries. Chimeric antigen receptors (CARs) and other antibody conjugates targeted to B7H3 are also described. The single-domain antibodies and conjugates thereof can be used for the diagnosis and treatment of B7H3 expressing solid tumors.

This invention relates to the field of biomedical technology, specifically to a snake-derived nanobody against KLH and its preparation and application. The invention involves immunizing the black-browed rat snake with KLH, extracting total RNA from its splenic lymphocytes, constructing a phage display VH gene library, and, after multiple rounds of screening, obtaining nanobody sequences that specifically bind to KLH, such as KLH-1 and KLH-2. This nanobody can be efficiently and solublely expressed and purified in *E. coli*, exhibiting characteristics of small molecular weight and high affinity. This invention further provides applications for this nanobody, particularly in the preparation of KLH-targeting detection reagents or drug delivery systems. This nanobody provides a core tool molecule for developing highly sensitive KLH-targeting detection reagents, efficient drug delivery systems, and novel vaccines.