280 results about "Titer" patented technology

The invention discloses preparation of a human CD56 immune phage display antibody library, which comprises the following steps: S1, animal immunization: expressing recombinant hCD56 protein in escherichia coli, then immunizing a Balb / c mouse, and detecting immune titer; s2, splenocyte preparation: extracting RNA (Ribonucleic Acid) of immune mouse spleen cells, and performing reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid); s3, preparation of an initial bacteriophage display library: amplifying a heavy chain variable region and a light chain variable region by using a PCR technology, and then assembling to form scFv; constructing scFv phagocytids, electrically transforming the scFv phagocytids into TG1 competent cells, and constructing a bacterial library; s4, constructing a phage single-chain library of the hCD56: infecting the bacterial library by using a helper phage M13K07, and constructing a phage single-chain antibody library; and S5, elutriation and screening of a phage library: specifically enriching a phage single-chain antibody library, screening positive clones from the phage single-chain antibody library, and determining a sequence. According to the invention, the human hCD56 antibody scFv sequence fragment can be screened out, and the research and development period is shortened.

The present invention belongs to the field of virus detection technology, and specifically relates to monoclonal antibodies with neutralizing activity against adenovirus type 5 and their uses. The monoclonal antibody 8B9 of the present invention can specifically recognize and neutralize adenovirus type 5, and the amino acid sequences of the heavy and light chain variable regions and their complementarity determining regions of the monoclonal antibody 8B9 are provided. The present invention also provides an in vitro neutralization activity assessment model based on HEK-293A cells. At a concentration as low as 3.3 μg / mL, the titer of the neutralizing activity against the virus stock solution after 6000-fold dilution is 7×10 4 TCID 50 / mL of virus has close to 100% neutralizing activity. After further humanization, it is expected to be developed into a neutralizing antibody drug for the treatment of diseases such as severe pneumonia caused by HADV-5 infection, filling the current lack of effective treatment options in this field and possessing significant scientific research value and clinical application prospects.

This invention discloses a monoclonal antibody against human hepatitis B e antigen and its application, relating to the field of hepatitis B detection technology. In the heavy chain variable region of the monoclonal antibody, the amino acid sequences of the complementarity-determining regions (CDR1-3) are as shown in SEQ ID NO. 1-3 or have at least 95% homology with the sequences shown in SEQ ID NO. 1-3; in the light chain variable region, the amino acid sequences of the complementarity-determining regions (CDR1 and CDR3) are as shown in SEQ ID NO. 4 and 5 or have at least 95% homology with the sequences shown in SEQ ID NO. 4 and 5, and the amino acid sequence of the complementarity-determining region (CDR2) is FAS. The monoclonal antibody provided by this invention can effectively recognize natural hepatitis B e antigen, and the titer of the supernatant from B cell culture can reach 1:10000, making it fully applicable to the detection and research of human hepatitis B e antigen. The chemiluminescent reagent prepared using the monoclonal antibody described in this invention has advantages such as high specificity, strong anti-interference ability, high detection sensitivity, and good stability, with almost no missed detections, and can replace imported reagents.

Methods and systems for monitoring and / or controlling production units used at various stages of biological product development and manufacturing are presented herein. In particular, in certain embodiments, the biological production monitoring and control techniques described herein utilize a mid-infrared (mid-IR) analyzer that is capable of obtaining mid-infrared spectral data from an aqueous sample in substantially real time. The techniques described herein can utilize this mid-infrared spectroscopic data to measure sample quality metrics, such as protein content, titer, secondary structure, aggregation, and the like, as well as viral and / or nucleic acid characteristics. Thus, sample quality metrics can be measured substantially in real time and / or continuously to assess, for example, the production quality of protein and gene therapeutic agents, and can be used by a control system to adjust and / or control process parameters to improve product quality, etc.

The invention discloses a surface antigen gene StSAG1 of Sarcocystis tenella, the nucleotide sequence of the surface antigen gene StSAG1 is as shown in SEQ ID NO: 1, and according to the analysis result of protein antigenicity and antigen epitope, the 686 bp-1175 bp segment of the StSAG1 gene is selected to construct a recombinant protein expression vector and engineering bacteria; the antigen disclosed by the invention has very strong antigenicity and specificity, and can be subjected to effective antigen-antibody reaction with an antibody generated in serum after sheep (Ovis aries) is infected with sarcosporidium tenella, so that diagnosis is realized; low-titer antibodies can be detected two weeks after infection, and the detection window period is obviously advanced; by adopting the ELISA detection method, a visual result can be directly obtained. The method breaks through the limitation of an infection window period, has the advantages of low cost, high sensitivity, simplicity and convenience in operation, no need of expensive instruments and analysis and the like, is suitable for field detection in primary laboratories and pastures, and is beneficial to industrial production and market popularization and application.

The present invention relates to the technical field of biomedicine. Specifically provided are a respiratory syncytial virus (RSV) Pre-F recombinant protein nanoparticle vaccine and a preparation method therefor. By performing amino acid mutation on an RSV Pre-F protein, and performing fusion expression on the mutated Pre-F protein and ferritin particles in eukaryotic cells, PreF-Ferritin fusion protein nanoparticles, in which eight Pre-F protein trimers are densely displayed on the surface of each ferritin particle, are obtained. By stabilizing and exposing antigen epitopes required to be displayed and disrupting or masking unrequired antigen epitopes, the PreF-Ferritin fusion protein nanoparticles effectively improve the immunogenicity and the production stability of the antigen. Experiments show that: when the PreF-Ferritin fusion protein is injected into mice, high-titer protective sera can be acquired, and the mice sera can yield relatively high neutralizing titers against RSV.

The invention provides a safrole hapten and application of the safrole hapten in detection of safrole through immunoassay. The structure of the safrole hapten is shown in the specification. The two safrole haptens provided by the invention have spacer arms with proper lengths, and after the safrole haptens are coupled with carrier protein, the haptens can be fully exposed, and recognition of the spacer arms can be avoided as much as possible. The safrole hapten is used for preparing an artificial antigen and an antibody, and the obtained safrole monoclonal antibody is high in titer, strong in specificity and high in affinity. An immunoassay method constructed on the basis of the safrole artificial antigen and antibody has high specificity and sensitivity, the lowest detection limit LOD is 0.09 ng / mL, the half inhibitory concentration IC50 is 2.05 ng / mL, the quantitative detection range is 0.28-14.89 ng / mL, no cross reaction exists on safrole analogues, and rapid qualitative and quantitative detection can be conducted on safrole in a sample.

The invention relates to the technical field of biological medicines, and discloses a PD-L1 K263 acetylation modified antibody, which is prepared from an immunogen containing a sequence as shown in SEQ ID NO.1. The PD-L1 K263 acetylation modified antibody is a PD-L1 K263 acetylation modified antibody. The invention also discloses a preparation method of the PD-L1 K263 acetylation modified antibody. The preparation method comprises the following steps: designing and synthesizing a polypeptide antigen, preparing an immunogen, immunizing animals, purifying the antibody and evaluating serum titer. The antibody provided by the invention can accurately recognize and combine with the PD-L1 protein subjected to K263 acetylation modification in the breast cancer, provides a specific tool for qualitative and quantitative detection of K263Ac-PD-L1 in the breast cancer, and provides a reliable molecular marker detection means for evaluating the curative effect of anti-PD-1 / PD-L1 immunotherapy; therefore, a biological preparation with specificity and practicability and a technical support are provided for accurate diagnosis, targeted therapy and curative effect monitoring of breast cancer.

The invention discloses a self-assembled ferritin nano antigen particle and application thereof in preparation of a duck hepatitis A vaccine. The VP1 part in the duck hepatitis A virus P1 protein is connected with the N end of a self-assembled ferritin nanoparticle subunit through a connecting sequence to obtain the fusion protein, and the amino acid sequence of the fusion protein is shown as SEQ ID NO.1; in order to improve the expression quantity or expression efficiency of the fusion protein in bombyx mori, a nucleotide sequence of a coding gene of the fusion protein is optimized and modified according to the codon preference of bombyx mori, and an optimized gene sequence with the nucleotide sequence shown as SEQ ID NO.2 is obtained; the fusion protein is subjected to single-site or multi-site mutation, the mutant with obviously improved titer is obtained through screening, and the soluble expression quantity of the mutant is obviously improved. According to the invention, a silkworm or insect cell eukaryotic expression system is used for expressing recombinant protein, and an expression product causes a neutralizing anti-duck hepatitis A virus antibody.

The invention provides a Garlu virus attenuated strain and application thereof, and relates to the field of virology and biological medicine. According to the invention, the screened Garlu virus attenuated strain is a natural attenuated strain derived from a tick sample, can grow in Vero cells, has the titer of more than or equal to 107 TCID50 / mL, does not have clinical pathogenicity to an immunodeficient host, and is safe; after the attenuated strain is used as an immunogen for pre-intervention, infection of a lethal virus strain can be avoided, the survival rate is increased to 100%, adverse effects on growth and life of experimental animals are avoided, particularly, a host can be stimulated to generate humoral immunity and cellular immunity at the same time, and the survival rate of the experimental animals is improved. And moreover, broad-spectrum cross-protection efficacy can be generated on Bandavirus lethal virus strains, and a foundation is laid for improvement of research on Bandavirus live vaccines such as the Colu virus and the like.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody 1E12 for specific detection of AAV6 and application of the monoclonal antibody 1E12. According to the monoclonal antibody 1E12 provided by the invention, a heavy chain variable region and a light chain variable region respectively comprise three complementary determining regions (CDR) corresponding to SEQ ID NO.1-3 and SEQ ID NO.4-6. The CDRs endow the antibody with high specificity and affinity to AAV6, the antibody can accurately recognize AAV6 virus without cross reaction with other serotypes, and the detection sensitivity reaches 4 * 10 < 6 > vg / ml level. The antibody is suitable for development of multiple detection tools such as a double-antibody sandwich ELISA kit, a test strip and an antibody chip, especially has excellent performance in a biotin-avidin amplification system, can be used for virus identification, titer determination and clinical dose evaluation of AAV6, and provides an effective means for standardized and high-throughput detection.

Disclosed herein are improved methods for quantifying monoclonal antibody (mAb) titer using Protein A affinity High-Performance Liquid Chromatography (HPLC). The methods address inaccuracies caused by mAb adsorption to sample vial surfaces by incorporating a surfactant into the mobile phase used for sample dilution. This approach enhances the accuracy, linearity, and robustness of the titer assay, making it suitable for process control and quality assessment in biopharmaceutical development and manufacturing. The methods can be applied as a platform approach for various mAb products.

The invention discloses a duck BRS3 polypeptide and a preparation method and application of a polyclonal antibody of the duck BRS3 polypeptide, and belongs to the field of biochemistry and molecular immunology. The amino acid sequence of the duck BRS3 polypeptide is as shown in SEQ ID NO. 3. A segment of polypeptide sequence with the length of 20 aa is screened out in an intracellular region of a duck BRS3 protein sequence, after artificial solid-phase synthesis, the polypeptide sequence is used as an immunizing antigen to immunize New Zealand white rabbits, and the duck BRS3 protein polyclonal antibody with the antiserum titer being 1: 25600 or above is obtained. The rabbit anti-duck BRS3 polyclonal antibody prepared by the invention can be specifically combined with the BRS3 protein in duck tissues, so that the blank in the field of detection and research of the duck BRS3 protein is filled, and a foundation is laid for research of functions of the BRS3 in duck bodies.

The invention belongs to the technical field of antibody preparation, and particularly relates to a recombinant rabbit monoclonal antibody for specifically recognizing type II grass carp reovirus VP56 protein and application of the recombinant rabbit monoclonal antibody. The amino acid sequence of a light chain variable region of the monoclonal antibody is as shown in SEQ ID NO.3, and the amino acid sequence of a heavy chain variable region of the monoclonal antibody is as shown in SEQ ID NO.1. The recombinant rabbit monoclonal antibody for resisting the II type grass carp reovirus VP56 protein prepared by the invention has relatively strong specificity and titer, can specifically recognize the virus VP56 protein in tissues and cells in a natural state, has high affinity for recognizing and combining the virus VP56 protein, has strong anti-interference capability, and can be used for preparing the recombinant rabbit monoclonal antibody for resisting the II type grass carp reovirus VP56 protein. The method is suitable for establishing a high-specificity, high-sensitivity and high-accuracy type II grass carp reovirus diagnosis technology.

A system for measuring an antibody concentration in a sample includes a polarizer holder holding a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from within a container to pass in a first direction. The second detection polarizer restricts fluorescence emitted from within the container to pass in a second direction. The second direction is perpendicular to the first direction. The system includes a detector for measuring fluorescence emitted in a first direction and fluorescence emitted in a second direction. The system rotates the polarizer holder in an alternating direction about an axis of rotation such that the first detection polarizer and the second detection polarizer are alternately placed in an optical path of fluorescence emitted from within the container.

The present application belongs to the technical field of antibody preparation, and belongs to a human CHI3L1 monoclonal antibody 1D7, a recombinant carrier, a recombinant plasmid and a method and application. The present application provides a monoclonal antibody 1D7 capable of specifically recognizing CHI3L1, wherein the monoclonal antibody 1D7 comprises a heavy chain and a light chain; a variable region of the heavy chain comprises three heavy chain complementarity determining regions, and the amino acid sequences of the three heavy chain complementarity determining regions are shown as SEQ ID NO. 1-SEQ ID NO. 3 or SEQ ID NO. 11-SEQ ID NO. 13; and a light chain variable region of the monoclonal antibody 1D7 comprises three light chain complementarity determining regions, and the amino acid sequences are shown as SEQ ID NO. 4-SEQ ID NO. 6. The monoclonal antibody provided in the present application has good titer, and the expression carrier and the recombinant cell for preparing the monoclonal antibody are easy to store and modify.

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 discloses a human intragastric factor protein polyclonal antibody and a preparation method thereof. The preparation method comprises the steps of artificial antigen design, animal immune optimization and antibody purification and verification. Accurate epitope design is achieved, structural interference of natural GIF is avoided through artificial polypeptide, and the cross reaction rate is lt; 5%; an efficient immune strategy is adopted, so that the immune period is shortened to 8-10 weeks, and the titer is increased by 2-3 times; low-cost production is realized, natural protein is replaced by the synthesized polypeptide, and the cost is reduced by 60%; the specificity is high, the cross reaction is reduced due to the design of the artificial antigen, and verification data supports specific recognition of natural GIF; large-scale production, polypeptide synthesis and a double-stage purification process are adopted, so that the consistency between batches is high (CV is less than or equal to 8%); the application range is wide, and diagnosis, pathological research and treatment monitoring scenes are covered.

This invention belongs to the fields of molecular biology and bioinformatics, specifically relating to a truncated bovine virus-encoded protein phage library, its construction method, and applications. This library covers all 162 known bovine viruses that host bovine subfamily viruses, containing 51,794 non-redundant peptide coding sequences of 56 amino acids each. These peptides are fused and displayed on the surface of the T7 phage capsid, with all coding sequences having a uniform total length of 200 bp. The library construction was achieved through sequence acquisition, truncation and redundancy removal, codon optimization and DNA synthesis, and in vitro ligation and packaging with T7 phage. The resulting library exhibits a positivity rate of 93% and a titer of 2.8 × 10⁻⁶. 10 PFU / mL. This invention features an optimized single-round high-throughput screening process adapted to clinical bovine serum samples, enabling efficient identification of bovine virus B-cell epitopes and providing tools and candidate targets for bovine virus vaccine development and diagnostic reagent development.

The invention discloses application of a DDIT3 gene as a target spot in regulation and control of bovine coronavirus replication. Experiments find that regulation and control of the expression level of the DDIT3 gene can influence the replication level of the BCoV, when the expression of the DDIT3 gene in a host cell is inhibited, the replication of the BCoV can be promoted and the titer of the virus can be improved, and when the DDIT3 is over-expressed on the host cell, the replication of the BCoV can be inhibited and the titer of the virus can be reduced. Experimental results show that the DDIT3 gene can be used as a novel target for developing BCoV antiviral drugs, and a tool and a material are provided for researching a molecular mechanism of the DDIT3 gene for regulating and controlling pathogenic microorganism replication in cells.

Provided are: a learned model, with which index data relating to the kinetics of contents in a container can be evaluated with high accuracy; an estimation device; and an estimation method. This learned model is trained using, as an explanatory variable, at least one of data relating to the shape of a stirrer provided in a container, data relating to the shape of a sparger provided in the container, data relating to the shape of the container, and data relating to the physical properties, the composition, the chemical properties, the biochemical properties or the concentration of contents accommodated in the container, and also using, as an objective variable, a mass transfer capacity coefficient (kLa), a stirring power (P / V), a respiratory rate, the concentration of bacterial bodies, the concentration of a product from the bacterial bodies and cells, or a titer.

Provided are a primer-probe combination for real-time quantitative PCR, a related kit, and a method for using primers to measure the titers of measles, mumps and varicella virus pathogens. The method is applicable to clinical and laboratory detection, is fast and convenient, and has high sensitivity.

The application relates to the field of biological medicine, and particularly discloses application of saliva TP17, TP47 and TP15 specific antibodies in preparation of an auxiliary diagnostic product for neurosyphilis. Based on specific antibody immunoglobulin G (IgG) of Treponema pallidum in saliva, combined detection of saliva TP17 / IgG, TP47 / IgG and TP15 / IgG outputs a risk value of neurosyphilis of a subject, or a four combined Logistic regression discriminant model is constructed by combining serum TRUST titers, and the subject is stratified into a low-risk or high-risk group based on a preset judgment threshold, so that the comprehensive recognition capability for neurosyphilis is improved. The saliva is used as a detection sample, and the application has the advantages of non-invasiveness, convenience, easy popularization, repeatable collection and the like, and can be used for early screening, auxiliary diagnosis and clinical path management of neurosyphilis.

The invention discloses a polyclonal antibody of panda Rbp7 protein as well as a preparation method and application thereof, and belongs to the technical field of antibodies. The first to nineteenth amino acid sequences of the panda Rbp7 protein are used as antigen polypeptides, and are specifically shown as SEQ ID No.1. After chemical synthesis or exogenous expression purification, a Balb / C mouse is immunized, and the polyclonal antibody is obtained through repeated immunization, serum titer detection and affinity chromatography purification. The titer of the antibody is not lower than 1 / 128000, the panda Rbp7 recombinant protein can be efficiently and specifically detected through WB and ELISA, the blank of a protein detection tool is filled, and reliable technical support is provided for research on the functional mechanism of the panda Rbp7 protein.

The invention relates to the field of bioengineering, in particular to a recombinant human herpes simplex virus as well as a construction method and application thereof. The recombinant human herpes simplex virus lacks US1, US2, US3, US4 and US5 virulence genes at the same time. The construction method comprises the following steps: constructing a donor plasmid, and constructing a cleavage plasmid; transfecting cells with the cleavage plasmids and the donor plasmids in proportion; and after the transfected cells are incubated, infecting with an HSV-1 virus, purifying, and verifying to obtain the recombinant virus. The donor plasmid constructed by the invention retains non-coding regions among US1, US2, US3, US4 and US5 genes, also retains initiation codons and termination codons of the US1, US2, US3, US4 and US5 genes, is added with a fluorescent tag sequence, and can knock out five virulence genes of HSV-1US1-US2-US3-US4-US5 at the same time. The neutralizing antibody titer result of the recombinant human herpes simplex virus is the same as that of a wild type, and a foundation is laid for vaccine preparation.

The present application relates to the antibody of phorbolylated isocitrate dehydrogenase 1 and its preparation method and purpose, and specifically provides a polyclonal antibody, the polyclonal antibody is obtained by immunizing animals with the antigen epitope peptide of IDH1 phorbolylated, the specific site of the antigen epitope peptide is phorbolylated, and high-titer antibodies can be prepared, the phorbolylated modification of IDH1 in clinical specimens can be recognized with high specificity, and the polyclonal antibody can be used for the research of cell / tissue specimen immunoblotting and immunohistochemical method and the diagnosis of IDH1 related diseases.

The invention discloses a polyclonal antibody targeting a human CPNE7 variable splicing isomer and application of the polyclonal antibody, and belongs to the technical field of biological medicine. The antigen epitope 'KYKQKRRSYKN' (SEQ ID NO.1) capable of being targeted by the polyclonal antibody provided by the invention is positioned in a common conserved region of all main isomers of CPNE7, and a variable splicing region is not involved. The titer of the obtained antibody serum is as high as 1: 32,000, which indicates that the antibody has extremely high immunoreactivity and sensitivity and can be used for detecting trace proteins. The antibody disclosed by the invention can be used for effectively identifying GST-CPNE7 fusion protein expressed by a prokaryotic system and CPNE7-FL and CPNE7-S protein overexpressed in an eukaryotic system (A549 cells), and the applicability of the antibody in various experimental systems is proved. Therefore, the blank in the prior art can be filled, and a core tool is provided for research of the CPNE7 in the fields of tumor biology, diagnosis and treatment.

The application provides a YKL-40 monoclonal antibody and a preparation method and application thereof, and relates to the technical field of biological medicines.The monoclonal antibody 10F1 provided by the application has good titer.The heavy chain variable region and the light chain variable region of the monoclonal antibody are respectively inserted into different basic vectors, and the two kinds of recombinant vectors are mixed and then co-transfected into basic cells, so that a recombinant cell capable of expressing the monoclonal antibody 10F1 is obtained.The monoclonal antibody vector obtained by the application is easy to preserve and easy to control the quality of the antibody production process; the monoclonal antibody vector can recognize YKL-40 protein and has biological activity.The monoclonal antibody provided by the application can be used for YKL-40 antigen detection and other scientific researches, and has very good application value and very important scientific research guiding significance.

The invention provides a method for predicting the ultraviolet inactivation effect of viruses in a liquid biological product. The method comprises the following steps: (1) detecting the absorbance value of a to-be-detected liquid biological product at a preset wavelength; (2) constructing a relation curve of the ultraviolet dose of the BSA / virus mixed solution and the decline degree of the virus titer; (3) constructing a relation curve of the ultraviolet dose of the SMZ working solution and the SMZ degradation degree; (4) constructing a relation curve of the operation parameters and the SMZ degradability; and (5) substituting preset operation parameters into the relation curve of the operation parameters and the SMZ degradability, substituting the obtained result into the relation curve of the ultraviolet dose and the SMZ degradability, substituting the obtained result into the relation curve of the ultraviolet dose and the decline of the indication virus titer, and judging the ultraviolet inactivation effect of the virus according to a predicted value of the decline of the indication virus titer. The method is accurate in prediction result, does not need to consume liquid biological products, and can be used for various virus inactivation effects under the condition that the ultraviolet dose cannot be accurately calculated.