123 results about "Antibody screening" patented technology

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody 4F6 for HPV16 type E7 protein detection and application. The heavy chain variable region CDR sequences and the light chain variable region CDR sequences of the antibody are SEQ ID NO.1-3 and SEQ ID NO.4-6 respectively, and the antibody can be specifically combined with HPV16 type E7 protein without cross reaction. A double-antibody sandwich ELISA detection system constructed based on the antibody is combined with a biotin-avidin amplification technology, the sensitivity reaches 100 pg / mL, and the antibody is suitable for rapid detection of the HPV16 type E7 protein in a cervical exfoliated cell sample. The invention further provides the HPV16 type E7 recombinant protein which is obtained through prokaryotic expression and has immunocompetence, and the HPV16 type E7 recombinant protein is applied to antibody screening and detection. Compared with nucleic acid detection, the method has a lower false positive rate, can be used for early screening of cervical cancer and precancerous lesions, and has a good application prospect.

The invention discloses a novel coronavirus neutralizing antibody, a detection kit and application of the novel coronavirus neutralizing antibody. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR12 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR12 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR12 to S-Trimer and RBD under 2-fold and 300-fold dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR12 has high affinity and dilution stability and is suitable for clinical large-dose administration.

The invention discloses a novel coronavirus neutralizing antibody, a detection kit and application of the novel coronavirus neutralizing antibody. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR75 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR75 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR75 to S-Trimer and RBD under 2-fold and 300-fold dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR75 has high affinity and dilution stability and is suitable for clinical large-dose administration.

The invention discloses a novel coronavirus neutralizing antibody GR46, a detection kit and application of the novel coronavirus neutralizing antibody GR46. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR46 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR46 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR46 to S-Trimer and RBD under 2-time and 300-time dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR46 has high affinity and dilution stability and is suitable for clinical large-dose administration.

The invention relates to a single bacterium separation method based on a digital micro-fluidic system, which comprises the following steps: injecting diluted bacterium liquid into a digital micro-fluidic chip to form bacterium liquid drops with 2 < N > power as electrode bases, and distributing 18-22 bacteria per 64 electrode bases in each bacterium liquid drop; the liquid drops with the same volume are controlled to move in the opposite direction, and half-and-half tearing is carried out; according to the mode, the liquid drops after half-and-half tearing are subjected to half-and-half tearing again, N rounds are carried out in total, and 2 < N > single-electrode liquid drops are formed; the single bacterium liquid drops are obtained. The separation method has the advantages that the separation efficiency is high, the separation speed is high, the separation flux is high, and the final separation success rate can reach 100%, and the single-bacterium separation method can be well applied to bacterium culture, polypeptide drug or antibody screening and genomics analysis in a digital microfluidic system.

This invention discloses a camel-derived CRP single-domain antibody, its preparation method, and its applications, relating to the biomedical field. The method involves enriching CRP antigen in alpaca leukocytes immunized with the antigen, followed by single-cell screening and sequencing. Through sequencing structural analysis, a recombinant plasmid of the camel-derived CRP antibody is constructed. This plasmid is cloned and transformed into eukaryotic cells to express related antibodies. Monoclonal antibodies with human CRP resistance are screened for expression. The VHH domain of the monoclonal antibody is cloned and expressed, and paired screening yields CRP-VHH monoclonal antibodies with antibody activity. This antibody can be used in ELISA kits, chemiluminescence kits, and latex kits, possessing the same functionality as existing kits. Furthermore, this antibody also has applications in the preparation of antitumor drugs.

The invention provides a disease marker epitope prediction and antibody screening method, which belongs to the technical field of disease markers, and comprises the following steps: firstly, establishing a training data set containing a known antigen-antibody compound structure and disease tissue expression data; and obtaining target protein sequence information through liquid chromatography-mass spectrometry analysis and carrying out sequence comparison. And then a deep convolutional neural network is utilized to extract sequence features, and surface exposure sites are identified by combining secondary structure prediction and solvent accessibility analysis. After the features are integrated with sequence evolution conservative properties, a prediction model is constructed by using a random forest classifier. The method comprises the following steps: carrying out molecular dynamics simulation on a prediction result, screening first 10% of candidate sequences through a comprehensive scoring function and K-means clustering, and finally determining an antigen epitope sequence with the strongest binding activity through verification of an antigen chip and a fluorescence labeled antibody system, so that the technical problem that specific antigen epitopes are difficult to accurately predict and recognize in the prior art is solved.

The invention discloses a method for high-purity sorting and capturing of rare cells. According to the method for high-purity sorting and capturing of the rare cells, the cells are spread firstly, then the positions of the target cells are searched and determined, then a probe is used for discharging a buffer solution outwards to blow away non-target cells around the target cells, and then fluid power is used for capturing the target cells without damage. Finally, the target cells can be released into a receiving container, and high-purity sorting and capturing of the rare cells are achieved. A device matched with the method is simple in structure and easy to operate, and the method has the advantages that repeated and massive dilution is not needed, the cell sample spreading speed is high, the target cell sorting purity is high, the omission rate is low, in-situ capture is achieved, and repeatability is good. The method is suitable for the fields of single cell screening and analysis, single cell culture, single cell drug screening, single molecule analysis, gene screening, sample purification, antibody screening, microbiological research and the like.

The invention provides an antibody screening method and system based on a differential evolution algorithm and an artificial intelligence model. The method comprises the following steps: S1, acquiring data of an antibody-antigen compound sequence and affinity; s2, encoding the antibody and antigen sequences by using a language model, and converting the encoded antibody and antigen sequences into a matrix M; s3, training a machine learning model by using the matrix M and the affinity parameters to obtain a trained optimal machine learning model; s4, predicting the antigen affinity of the antibody by utilizing the trained optimal machine learning model; and S5, carrying out affinity optimization on the antibody sequence based on a differential evolution algorithm, and carrying out further filtration by predicting the druggability of the antibody so as to screen the antibody with high affinity and good drug properties.

The invention provides an antibody screening method for flow cytometry, and belongs to the technical field of flow cytometry. Antibody purity analysis and concentration determination are carried out through high performance liquid chromatography; the binding characteristic of the antibody and the target cell is researched by adopting an equal-ratio dilution method; analyzing the combined signal by using an improved Hilbert-Huang decomposition algorithm; evaluating the combination stability through covariance matrix decomposition; collecting cell event data by using a flow cytometer; constructing a fluorescence intensity contribution matrix and performing signal optimization; calculating a signal stability contribution value to determine an effective detection area; a 0-1 knapsack optimization method is adopted to carry out multiple detection combination optimization; finally, repeatability verification is carried out through a bootstrap resampling method, the stable-performance antibody for flow detection is screened out, and the technical problems that in the prior art, the signal stability of antibody screening for flow detection is poor, and specificity is insufficient are solved.

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 relates to an erythrocyte specific antibody screening test kit expressing a single antigen from a target erythrocyte (RBC) antigen (RhD and RhCE as 2 different antigens derived from RBC and 45 or more antigens in total including some variants of these two antigens) and a preparation method of this test kit. Said antigens are transgenically produced and provide the clearest results compared to traditional methods of antibody screening. Field-targeted mutation process is applied during the preparation of the erythrocyte-specific antibody screening test kit subject to the invention.

The present invention provides a method for preparing a reagent for microfluidic antibody screening, which is carried out according to the following steps: step 1, preparing corresponding reagents; step 2, pushing B cell reagents; step 3, mixing B cells with a trace amount of serum; step 4, generating screening reagents and agglutination samples; step 5, observing the agglutination state and selecting the corresponding screening reagents; step 6, extracting the screening reagents by using microfluidic technology and performing cloning culture. The preparation method and device can simultaneously mix the multi-component B cell reagents with the serum reagents on the top through the pushing mechanism at the front end, thereby providing multiple agglutination samples and screening reagents at the same time, improving the success rate of antibody production, and directly corresponding each agglutination sample to one of the screening reagents, so as to accurately judge the internal antibody production state of each screening sample, and avoid the situation where the antibody components of the agglutination sample and the screening reagent are inconsistent.

Herein is reported a method for preparing a recombinant host cell library expressing bispecific antibodies using targeted integration into a host cell comprising an exogenous nucleotide sequence integrated at a site within a locus of the genome of the host cell, wherein the exogenous nucleotide sequence comprises a first and a second recombination recognition sequence flanking at least one first selection marker, and a third recombination recognition sequence located between the first and the second, recombination recognition sequence, and all the recombination recognition sequences are different whereby the host cell is transfected with a library of first vectors each comprising two recombination recognition sequences matching the first and the third recombination recognition sequence on the integrated exogenous nucleotide sequence and flanking two exogenous nucleotide sequences and at least one second selection marker, and a library of second vectors each comprising two recombination recognition sequences matching the second and the third recombination recognition sequence on the integrated exogenous nucleotide sequence and flanking at least two further exogenous nucleotide sequences; wherein one of the four exogenous nucleotide sequences encode a first light chain, one a second light chain, one a first heavy chain and one a second heavy chain of the bispecific antibody.

The invention relates to the field of antibodies, and particularly provides a high-affinity PADI2 antibody and a preparation method thereof. According to the invention, eight high-affinity PADI2 antibodies are obtained by screening through an antibody pair screening experiment, and the antibodies can be specifically combined with PADI2 protein and have high affinity. Experimental results show that the antibody disclosed by the invention shows excellent binding capacity in ELISA (Enzyme-Linked Immunosorbent Assay), Western blot and cell experiments, and can be used for detecting, diagnosing and treating PADI2 related diseases.

The application discloses a CRP camel-derived single-domain antibody and a preparation method and application thereof, relates to the field of biological medicines, and is characterized in that: white blood cells of a llama immunized with a CRP antigen are enriched, and then single-cell screening and sequencing are performed. Through sequencing structure analysis, a camel-derived CRP antibody recombinant plasmid is constructed, a related antibody is expressed in a eukaryotic cell through cloning transformation, a single antibody with human CRP resistance is screened, a VHH domain of the single antibody is cloned and expressed, and a single antibody with CRP-VHH antibody activity is obtained through pairing screening. The antibody can be applied to an ELISA kit, a chemiluminescence kit and a latex kit, has the same functions as existing kits, and still has the use in the preparation of an anti-tumor drug.

The invention provides a heterochromatin protein HP1 and histone H2A-H2B assembled protein complex, a preparation method and an antinuclear antibody screening kit, the protein complex is formed by combining the histone H2A-H2B and an HP1 protein C-terminal truncation, and the selection region of the HP1 protein C-terminal truncation is 109-175 AA; according to the invention, the HP1-CSD and the histone H2A-H2B are assembled in vitro, the C-terminal truncated body of the heterochromatin protein HP1 or the HP1 protein can be directly combined with the histone H2A-H2B, and more theoretical references are provided for subsequent research on a molecular mechanism formed by the HP1-mediated heterochromatin. In addition, the invention provides an antinuclear antibody screening kit, the kit comprises the protein complex, and the reagent stability is improved.

The invention discloses a real-time detection system and a monitoring and early warning method for African swine fever virus. The system comprises a latex microsphere labeled immunochromatography antibody detection card, a portable MINI-PCR instrument and a freeze-dried microsphere type fluorescent PCR detection reagent, the detection card takes the recombinant ASFV p30 protein as a target spot, antibody detection is completed within 10 minutes, the lowest detection limit is 1: 1024, and the specificity is greater than 99%; the PCR reagent adopts a double-primer double-probe design, and the lowest detection limit is 1 copy / microliter. According to the invention, through a collaborative process of antibody screening, nucleic acid detection and result evaluation, ASFV total infection cycle detection is realized, epidemic situation discovery time is advanced to 1-3 days after infection, and early warning response time is shortened by more than 60%; the system is simple to operate and low in cost, does not need professionals and laboratories, can be deployed in basic-level scenes such as pig farms and slaughter houses, effectively solves pain points of an existing detection system, and provides key technical support for prevention and control of African swine fever.

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 invention relates to a fully humanized single-domain antibody for resisting Glypican-3 and an application of the fully humanized single-domain antibody. The antibody comprises three heavy chain complementary determining regions (CDRs), and the amino acid sequence of the CDRs is the 31st-35th amino acid sequence of SEQ ID NO: 1; the amino acid sequence of the CDR2 is the 50th-66th amino acid sequence of SEQ ID NO: 1; the amino acid sequence of the CDR3 is the 99-107th amino acid sequence of SEQ ID NO: 1. According to the invention, a bacteriophage display technology is utilized, antibody screening is carried out through targeting human GPC3 protein, and one anti-human GPC3 specific fully-humanized monoclonal single-domain antibody is obtained; the antibody can be specifically combined with human GPC3, can effectively inhibit activation of a Wnt signal, provides an effective alternative antibody drug for HCC treatment with GPC3 as a target, and has a potential clinical application prospect.

The application discloses an antibody for detecting a novel duck reovirus (NDRV), and relates to the field of animal disease prevention and treatment.The antibody comprises a heavy chain and a light chain, wherein the amino acid sequence of the heavy chain is shown as SEQ ID NO.2, and the amino acid sequence of the light chain is shown as SEQ ID NO.4.The novel duck reovirus sigma C protein antibody can be used for directly and rapidly detecting the NDRV sigma C protein in a quick, sensitive and accurate manner when the novel duck reovirus is detected.The antibody can be used for early diagnosis of the NDRV, epidemiological investigation, and antibody screening in vaccine development, and provides an effective tool for preventing and controlling the novel duck reovirus infection.

The invention discloses a risk assessment system for difficulty in cross matching of blood, and relates to the technical field of clinical medical examination and blood transfusion management. The system comprises a data input module, a risk calculation module, a risk layering module and a result output and suggestion module. The data input module obtains key information of a patient, wherein the key information comprises an erythrocyte irregular antibody screening result, multiple myeloma diagnosis, hematology department medical history, age of more than or equal to 60 years, pregnancy-related disease state and blood type identification difficulty; the risk calculation module adopts a preset scoring algorithm to endow each factor with a weight and calculates a total risk score; the risk layering module divides the patients into three levels of low risk, medium risk and high risk; and the result output module provides differentiated clinical management paths corresponding to the risk levels. According to the method, early warning of difficulty in cross matching of blood is achieved through quantitative evaluation, blood transfusion process efficiency can be optimized, clinical risks can be reduced, and the method has good prediction performance (AUC = 0.819) and a high negative prediction value (93.7%).

A homogeneous method for detecting the nucleocapsid protein of the new coronavirus based on a nano-antibody-monoclonal antibody sandwich relates to the field of antigen detection. The present invention prepares a variety of novel coronavirus nucleocapsid protein nano-antibodies as solid-phase antibodies, nucleocapsid protein monoclonal antibodies as detection antibodies, and novel coronavirus N protein as detection targets. A group of antibody pairs with excellent detection performance are screened out by enzyme-linked immunosorbent assay, with a minimum detection line of 0.8 ng / mL. In addition, a one-step activation method couples nano-antibodies and monoclonal antibodies to the surfaces of quantum dot microspheres and magnetic nano-microspheres, respectively, and successfully prepares two probes targeting the nucleocapsid protein of the new coronavirus. It also constructs a magnetic separation immunohomogeneous method for rapid detection of the nucleocapsid protein of the new coronavirus, with a minimum detection line of 0.15 ng / mL. The present invention is simple to operate, low in cost, fast in reaction speed, can realize in situ detection, and has high efficiency in paired antibody screening. It has good application prospects in the rapid detection of novel coronaviruses.

The invention relates to the technical field of medicine production equipment, in particular to an antibody screening and filtering device which comprises a base, a filtering box, a feeding port, a filtering mechanism, a driving mechanism, a cleaning mechanism and a backwashing mechanism, the filtering box is fixedly installed on the base, the feeding port is fixedly installed on the filtering box, and the filtering mechanism is fixedly installed on the filtering box; the filtering mechanism is used for filtering antibodies in the filtering box, the cleaning mechanism and the backwashing mechanism are fixedly mounted on the driving mechanism, the cleaning mechanism is used for cleaning the filtering mechanism, the backwashing mechanism is used for backwashing the filtering mechanism, and the driving mechanism can be mounted on the filtering box in a sliding manner. According to the antibody screening and filtering device, through the arrangement of the filtering mechanism and the driving mechanism, the position of the filter membrane can be adjusted, so that subsequent antibodies can be filtered on the clean filter membrane surface, and the use effect and the working efficiency of the antibody screening and filtering device are improved.

The invention belongs to the technical field of bioengineering, and particularly relates to a preparation method and application of a calprotectin S100A8 / A9 dimer mouse monoclonal antibody, the monoclonal antibody comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises any one of amino acid sequences as shown in SEQ ID NO.22-24; the heavy chain variable region comprises any one of amino acid sequences as shown in SEQ ID NO.22-24; the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO. 28-30. According to the present invention, the S100A8 / A9 heterodimer protein with biological activity and immunostimulation function is prepared, the protein is used to immunize mice, the specific gene engineering antibody of the S100A8 / A9 heterodimer protein is prepared, the antibody pair with high sensitivity is screened, and the calprotectin S100A8 / A9 double-antibody sandwich enzyme-linked immunosorbent assay method is preliminarily developed;

1. The name of the design product: full-automatic blood type analyzer. 2. The use of the design product: using column agglutination method, together with the matching detection reagent, for ABO system blood type typing, Rh system blood type typing, irregular antibody screening, cross matching and direct anti-human globulin test on human blood samples in clinical application. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective view.

The application discloses a kind of based on HIV envelope protein chimeric exosome and its preparation method and application, belong to biological medicine technical field.The application first constructs the cell line of stable expression HIV envelope protein Env, obtains engineered exosome from cell culture supernatant separation and purification;The exosome is used as immunogen combined with adjuvant immunization experimental animal, and high-efficiency induction specific humoral immune response;Again, obtain Env antigen specificity single B cell by flow cytometry sorting, obtain antibody variable region gene by single cell lysis, reverse transcription and nest PCR amplification, cloning to expression vector and expressing in mammalian cell, finally, HIV specific neutralizing antibody is screened by binding activity, affinity and neutralizing activity;The application is combined with single B cell antibody screening technology by embedding HIV Env antigen in the form of membrane combination in exosome surface, and realizes the synergistic optimization of antigen delivery and antibody screening process.

The present application relates to the technical field of biotechnology, in particular to a monoclonal antibody against A29L protein of monkeypox virus and application thereof. The present application prepares the monoclonal antibody against A29L protein of monkeypox virus, and through optimization of antibody screening and expression technology, the antibody with high affinity and high specificity is screened, and then a new tool is provided for rapid detection, effective neutralization and treatment of monkeypox virus, and an important immunological basis is provided for vaccine research and development and treatment scheme design of monkeypox virus.

The present invention relates to a method of detecting intermolecular interaction kinetics, the method comprising a first molecule and a second molecule to be analyzed, dispersing the first molecule in a solution as a detection solution, attaching the second molecule to a biosensor and being freely translatable over the surface of the sensor; the biosensor comprises a tethered bilayer phospholipid membrane and a solid conductive surface, a lower layer membrane of the tethered bilayer phospholipid membrane is fixed, and an upper layer membrane of the tethered bilayer phospholipid membrane can freely translate; the phospholipid membrane contains ion channel protein; and contacting the detection solution with the biosensor, detecting the conductivity change of the tethered bilayer phospholipid membrane, and analyzing the interaction kinetics of the first molecule and the second molecule. The method for detecting intermolecular interaction kinetics based on tethered bilayer phospholipid membrane and ion channel switch technology development can realize accurate quantitative detection in a wide concentration range, does not need sample flow, and is suitable for the fields of antibody screening, drug screening and life science.

The present invention discloses a method for high-purity sorting and capturing of rare cells. The method for high-purity sorting and capturing of rare cells in the present invention first spreads the cells, then locates and determines the position of the target cells, then uses a probe to expel the buffer outward to disperse the non-target cells around the target cells, and then uses hydrodynamic force to capture the target cells without damage. Finally, the target cells can be released into a receiving container to achieve high-purity sorting and capturing of rare cells. The device used in the method of the present invention has a simple structure and is easy to operate. The method has the advantages of not requiring repeated large-scale dilution, fast spreading speed of cell samples, high sorting purity of target cells, low omission rate, in-situ capture, and good repeatability, and is applicable to fields such as single-cell screening and analysis, single-cell culture, single-cell drug screening, single-molecule analysis, gene screening, sample purification, antibody screening, and microbial research.