699 results about "Antiserum" patented technology

The present invention is directed to the cloning, sequencing and expression of homologous immunoreactive 28-kDa protein genes, p28-1, -2, -3, -5, -6, -7, -9, from a polymorphic multiple gene family of Ehrlichia canis. Further disclosed is a multigene locus encoding all nine homologous 28-kDa protein genes of Ehrlichia canis. Recombinant Ehrlichia canis 28-kDa proteins react with convalescent phase antiserum from an E. canis-infected dog, and may be useful in the development of vaccines and serodiagnostics that are particularly effective for disease prevention and serodiagnosis.

Non-infectious, retrovirus-like particles comprise an assembly of an env gene product, a pol gene product and a gag gene product contain an antigenic marker which is non-retroviral or non-HIV retroviral. In one embodiment, the marker comprises an amino acid sequence containing an epitope inserted into the gag gene product at an antigenically-active insertion site. In another embodiment, the marker comprises an antigenic anchor sequence operatively connected to the env gene product replacing endogenous anchoring function. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in in vivo administration including to humans and in diagnosis. The presence of the antigenic marker enables recognition that antiserum containing anti-retroviral antibodies has been generated by exposure to the non-infectious retrovirus-like particles by testing for antibodies specific to the antigenic marker.

The invention discloses a method of preparing a bombyx mori silk fibroin specific antibody by utilizing a characteristic polypeptide. The method comprises the following steps: synthesizing a "CGAGAGSGAGAGS" polypeptide sequence by utilizing an Fmoc method, coupling the polypeptide with keyhole limpet hemocyanin (KLH) through the cysteine on the N terminus of the polypeptide so as to obtain a complete antigen; diluting the complete antigen with normal saline, mixing the diluted complete antigen with a complete Freund's adjuvant, adding streptomycin and penicillin to carry out an emulsion treatment so as to obtain a primary immunized antigen emulsion, subjecting a rabbit to a primary immunization by using the primary immunized antigen emulsion, then subjecting the rabbit to a strengthened immunization, wherein the strengthened immunization uses a strengthened immunized antigen emulsion, which is prepared by the following steps: mixing the diluted complete antigen with an incomplete Freund's adjuvant, and then adding streptomycin and penicillin to carry out an emulsion treatment so as to obtain the target product; collecting the blood of the immunized rabbit, when the antibody titer in the rabbit blood sample reaches 1 / 10000; making the blood blocks fully contract to completely separate out the antiserum, then collecting the antiserum, and subjecting the antiserum to a centrifugation treatment so as to obtain a supernate.

The invention belongs to the technical field of biological engineering and in particular is a heat-induced fusion expression recombinant plasmid of truncated GST protein and a process for preparation. The invention selects utility truncated GST188 protein which is composed of 188aa as a carrier, a pXXGST-1 recombinant plasmid which expresses short peptide fusion protein in an escherichia coli heat-induced expression system and a heat-induced type pXXGST-2 recombinant plasmid which is used to express comparison to realize the aim of synthesizing short peptide creatures, which is especially provided for scanning, drawing, positioning antigen linear epitopes and identifying epitope motif. Experiments of the invention prove the adaptability and the applicability of the GST188 core protein which is used as the short peptide creatures expression vector, in antigen epitope scanning identification, and in particular when anti- recombinant target protein antiserum is used to indentify the epitope motif.

Non-infectious, retrovirus-like particles comprise an assembly of an env gene product, a pol gene product and a gag gene product contain an antigenic marker which is non-retroviral or non-HIV retroviral. In one embodiment, the marker comprises an amino acid sequence containing an epitope inserted into the gag gene product at an antigenically-active insertion site. In another embodiment, the marker comprises an antigenic anchor sequence operatively connected to the env gene product replacing endogenous anchoring function. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in in vivo administration including to humans and in diagnosis. The presence of the antigenic marker enables recognition that antiserum containing anti-retroviral antibodies has been generated by exposure to the non-infectious retrovirus-like particles by testing for antibodies specific to the antigenic marker.

Compositions comprising a purified and / or isolated antibody, humanized antibodies, precipitates and anti-sera that specifically bind to or are otherwise directed against ROR1 protein. The compositions may be used for detecting ROR1 in a sample from a subject that is suspected or known to contain cancer cells. The ROR1 antibodies are especially useful in identifying and treating lymphomas and ademocarcinomas. Vaccines and related methods for protecting a subject against diseases that involve expression of ROR1 are also provided, as are human anti-sera effective in abrogating interactions between Wnt5a protein and ROR1 that contribute to the survival of certain cancer cells, such as CLL cells.

The invention relates to a reagent for determining APOE in blood serum, aiming to provide a stable reagent which has simple operation, high accuracy, good repeatability and strong anti-jamming capability, uses excellent new-born calf serum to replace human blood serum as substrate preparation and uses liquid blood serum type constant value calibration solution to detect apolipoprotein E in combination, wherein the sample does not need thinning dilution. The reagent is characterized in that the apolipoprotein E testing reagent comprises: a. a reagent R1 comprising buffer solution, a surface active agent, an electrolyte, a polymer accelerant, a reaction promoter, a stabilizer, a proper amount of preservative, and the balance of purified water; b. a reagent R2, wherein a certain amount of the reagent R1 is added to anti-human-APOE antiserum and antioxidant with the vacuum ratio of 20-50%; and c. constant value calibration solution comprising buffer solution, APOE antigen, antioxidant, synergist, stabilizer, proper amount of preservative, and the balance of purified water.

Non-infectious, retrovirus-like particles comprise an assembly of an env gene product, a pol gene product and a gag gene product contain an antigenic marker which is non-retroviral or non-HIV retroviral. In one embodiment, the marker comprises an amino acid sequence containing an epitope inserted into the gag gene product at an antigenically-active insertion site. In another embodiment, the marker comprises an antigenic anchor sequence operatively connected to the env gene product replacing endogenous anchoring function. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in in vivo administration including to humans and in diagnosis. The presence of the antigenic marker enables recognition that antiserum containing anti-retroviral antibodies has been generated by exposure to the non-infectious retrovirus-like particles by testing for antibodies specific to the antigenic marker.

In order to accurately and reliably quantitate HLE on the plasma membranes of the lymphocytes and mononuclear phagocytes, a test sample containing the lymphocytes and mononuclear phagocytes is initially treated with a first antiserum specific for CD4 receptors on the plasma membrane or with a second antiserum specific for chemokine receptors on the plasma membrane. Once the CD4 or chemokine receptors have been rendered non-reactive (competitive) relative to the HLE receptors (also “binding sites”) on the plasma membrane, the test sample is contacted with an immunoreagent specific for interaction with one or more of the HLE receptors on the plasma membranes of the lymphocytes and mononuclear phagocytes. The immunoreagent forms a complex with the HLE binding sites and produces a characteristic physical change in the lymphocytes and mononuclear phagocytes that can be monitored by any one of a number of standard techniques, (e.g., confocal laser scanning microscopy and flow cytometry).

The invention discloses an E.tenella protein disulfide linkage isomerase gene EtPDI (Clone ID is BW1-E06,and the Genbank accession number of is EF552214). The gene is connected with a procaryon expression vector pGEX-4T-2; a procaryon expression recombination plasmid pGEX-4T-EtPDI is constructed and is expressed in a colibacillus system; and most of the expressed recombining protein exists in a soluble form. The recombining protein 4T-EtPDI is purified to carry out SPS-PAGE and is transferred to a PVDF film; and antiserum of E.tenella oocyst oral immunized chicken is used as first resistance and goat anti-chicken IgG is used as second resistance to carry out Western-blot analysis, thereby indicating that the gene has certain antigen. The gene is used for preparing an anti-chicken coccidiosis drug and an anti-chicken coccidiosis vaccine.

The invention relates to polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine copolymer, a preparation method thereof and application thereof. The preparation method comprises the step of directly condensing carboxyl in the polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine segmented copolymer with amino in polyethyleneimine to form the graft copolymer. The copolymer is a polycation gene carrier, integrates the advantages of polyethylene glycol, Makrolan and polyethyleneimine, and has high transfection efficiency, wherein the highest transfection efficiency to the medium luciferase plasmid of african green monkey kidney cell is 14 times of that of American Invitrogen biological company transfection reagent Lipofetamine<TM>2000, can effectively antagonize the inhibiting effect of blood serum to the transfection, and has less cell toxicity, wherein consistency thereof is not higher than 200 microgramme / ml and cell survival rate is more than 80%.

The invention discloses a method of preparing a bombyx mori silk fibroin specific antibody by utilizing a characteristic dodecapeptide. The method comprises the following steps: synthesizing a polypeptide with a "CGYGAGAGAGYGA" sequence, coupling the polypeptide with keyhole limpet hemocyanin (KLH) so as to obtain a complete antigen; diluting the complete antigen with normal saline, mixing the diluted complete antigen with a complete Freund's adjuvant, carrying out an emulsion treatment so as to obtain primary immunized antigen emulsion, subjecting a rabbit to a primary immunization by using the primary immunized antigen emulsion, then subjecting the rabbit to a strengthened immunization, wherein the strengthened immunization uses a strengthened immunized antigen emulsion, which is prepared by the following steps: mixing the diluted complete antigen with an incomplete Freund's adjuvant, and then carrying out an emulsion treatment so as to obtain the target product; collecting the blood of the immunized rabbit, when the antiserum titer of rabbit arrives at 1 / 10000; making the blood blocks fully contract to completely separate out the antiserum, then collecting the antiserum, and subjecting the antiserum to a centrifugation treatment so as to obtain a supernate. The antibody prepared by the invention has a strong specificity, and can be used for detection and analysis of silk fibroin in textile, and the like.

The invention provides a kit for detecting serum amyloid protein and application thereof. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is prepared from a buffer solution, inorganic salt, a surfactant, a preservative, a stabilizer and interference elimination protein; the reagent R2 is prepared from the buffer solution, the inorganic salt, the surfactant, the preservative, the stabilizer and a polystyrene latex particle mixture; the polystyrene latex particle mixture is cross-linked with an SAA (Serum Amyloid A) antibody; the polystyrene latex particle mixture is a mixture of large-diameter polystyrene latex particles and small-diameter polystyrene latex particles. The kit disclosed by the invention is based on PETIA (Particle-enhanced Turbidimetric Immunoassay) and can be generally used for analysis of all kinds of full-automatic biochemical analyzer; during use, the required determining time is short, the specificity is high, the precision degree is high, and the accuracy degree is high.

A synthetic method of a general artificial antigen of phthalate plasticizers for immunodetection belongs to the field of bio-chemical engineering technology. The synthetic method of the present invention comprises the following steps of carrying out an esterification reaction between phthalic acid and 6-(fluorenyl methoxy carbonyl acyl-amino)-1-hexanol, removing fluorene methoxy carbonyl acyl protecting groups to form a hapten, and coupling with amino from carrier protein to obtain the general artificial antigen of phthalate plasticizers. Experimental results disclose that titer of antiserum obtained by immunizing animals by the antigen of the invention is up to 160000;the 50% inhibiting concentration IC50 for DBP, DEHP and DINP is less than 500 ng / ml; and the generated antibodies have good generality and high sensitivity. The antigen or antibodies of the invention can be used for the establishment of enzyme-linked immunosorbent assay and colloidal gold test paper rapid detection methods, thus can be used for rapid detection of residues of phthalic acid ester plasticizers in food, and have broad application prospects.

The invention discloses a recombinant human pepsinogen II isozyme chimeric protein, and a preparation method and applications thereof. The preparation method comprises the steps as follows: by taking gene sequences of two isozymes of recombinant human pepsinogen II as a template, carrying out PCR (polymerase chain reaction) splicing to obtain a chimeric protein coding gene sequence, constructing recombinant expression plasmids, transforming the screened positive clone plasmids into expression host cells, screening an efficiently-expressed recombinant chimeric protein strain, carrying out enlargement culture on the cells and inducing expression chimeric protein, and purifying to obtain the recombinant human pepsinogen II isozyme chimeric protein. The invention further discloses the applications of the recombinant human pepsinogen II isozyme chimeric protein in preparing monoclonal antibodies and multiresistant serums or pepsinogen II kit calibration products. A great amount of stably expressed recombinant human pepsinogen II isozyme chimeric protein can be produced by utilizing a genetic engineering technology, and one protein has two chimeric protein sequences of the human pepsinogen II.

The present invention discloses one kind of retinol conjugated protein and its preparation process and specific antibody. The present invention expresses retinol conjugated protein effectively for the first time, and the expressed retinol conjugated protein has property near to that of natural retinol conjugated protein (RBP4). Immunizing animal with the expressed retinol conjugated protein can obtain antibody capable of recognizing natural retinol conjugated protein in human body sensitively. The present invention fills in a gap in RBP4 antiserum.

This invention relates to hydrochloric acid CLB enzyme immunity test reagent box and its test method adopting ELISA competition to test CLB and using CLB and OA coupler as CLB antigen immunity rabbits to small mouses to get multiclonal containing CLB antibody or monoclonal CLB antiserum then to be put on enzyme specimen board after separation, purification and dilution to be warm cultivated, washed, added with sample diluent solution and enzyme specimen antigen to be reacted, washed and added with enzyme substrate to display color. It is an easy test method with simple pretreatment especially for testing remaining CLB of meat food.

The invention relates to a polyclonal antibody for detecting the residue and the degradation of a CP4-EPSPS protein in a processed food and application thereof. An animal is immunized by using a CP4-EPSPS protein peptide segment of a coupled keyhole limpet hemocyanin (KHL) to obtain antiserum, namely the antibody of the invention. The application comprises a Western blotting and a colloidal gold test strip and has the characteristics of simplicity, practicability, high sensitivity and the like, wherein the Western blotting is used for detecting the residue and the degradation of the CP4-EPSPS protein in food; and test strip technology is used for the rapid detection of the CP4-EPSPS protein in the food. The invention provides a high-practicability method for the safety detection of a genetically modified food, and has a very high practical application value.

The present invention provides a non-human animal-derived antiserum and / or non-human animal-derived immunoglobulin against the 2019-nCOV (SARS-CoV-2). The invention is intended to be able to be used in the treatment and prevention of novel coronavirus pneumonia.

The invention relates to a human papilloma virus capsid protein L1 (HPVL1) peptide and preparation and application thereof. The HPVL1 peptide is a peptide segment of which the amino acid of the sequence is N end-Cys Thr Leu Thr Ala Asp Val Met listed Thr Tyr Ile His-C end, or is a peptide segment comprising the amino acid of the sequence and with a length less than 50 amino acids. In the invention, the peptide which is a very conservative peptide segment located on the surface of protein is sieved out by performing epitope forecasting and multi-sequencing comparison on the L1 proteins of 18 low-risk types and high-risk type HPV subtypes (6, 11, 16, 18, 52, 58 and the like); the antibody generated from induction can react with multiple types of HPVL1, and can be used for the detection of the multiple types of HPVL1, in particular to the application of precancerous lesions detection of human cervical carcinoma; and the antibody aiming at the peptide segment also can be used for purifying and preparing the multiple types of HPVL1. In addition, the invention also provides the applications of the peptide segment used as multiple types of HPV antiserum and detected matters of monoclonal antibody.

The invention discloses a synthesis method of a specific salbutamol artificial antigen, belonging to the technical field of biological chemical engineering. The synthesis method disclosed by the invention comprises the following steps of: activating salbutamol through a formaldehyde solution, and connecting 6-aminocaproic acid in the ortho-position of a phenolic hydroxyl group to obtain a salbutamol hapten; and coupling a carboxyl group on the salbutamol hapten with an amino group on a carrier protein to obtain the salbutamol artificial antigen. The synthesis method disclosed by the invention can make up for the insufficiencies and defects of the existing salbutamol antigen synthesis technologies, the salbutamol artificial antigen with high specificity is obtained, the specificity of a produced antibody is high, and the sensitivity is high; and experimental results show that the antiserum titre of an animal immunized by using the salbutamol artificial antigen disclosed by the invention can achieve 80000, the detection limit is 0.5ng / mL, and the half-inhibitory concentration IC50 is 5ng / mL. The antigen or the antibody disclosed by the invention can be used for establishing an enzyme-linked immunosorbent analytical method and a colloidal gold test strip rapid assay method so as to rapidly detect the residues of the salbutamol in a food and further realize broad application prospects.

The invention discloses half antigen and antigen of tenuazonic acid and a preparation method and application thereof. The antigen can serve as a compound of the tenuazonic acid half antigen and is provided with the structure as the formula I. The antigen of the tenuazonic acid is obtained by mixing TeAH and carrier protein, dissolving carrier protein into phosphate buffer solution (PBS), droppingglutaraldehyde solution, conducting stirring and reaction and then conducting dialysis through physiological saline, or the ntigen of the tenuazonic acid is obtained by dissolving TeAHGA into N, N-dimethylformamide, adding bicyclocaproyl carbodiimide and N-hydroxy succinimide into the mixture, stirring the liquid, conducting centrifugation to obtain supernatant as A liquid, dissolving carrier protein in the PBS, stirring and dissolving the PBS to obtain B liquid, dropping the A liquid into the B liquid for reaction and conducting dialysis through physiological saline. The antiserum valence obtained by antigen immune animals can reach 1: 3.2*104, the linear range of TeAH ranges from 0.130ng / mL to 8.789ng / mL(IC20-IC80), the half inhibition concentration is 1.095ng / mL, and the produced antigen is high in specificity, high in sensitivity and high in accuracy. Formula I.

Relating to the technical field of virus detection in genetic engineering, the invention provides a method for rapid and accurate detection of tomato yellow leaf curl virus (TYLCV). The method comprises: cloning a TYLCV-CP gene segment from the total DNA of a diseased leaf, connecting the TYLCV-CP gene segment with pET-32a and transforming a recipient bacterium, conducting inducible expression to a positive bacterial colony and purifying the target protein; preparing antiserum with an immunized rabbit, labeling a purified antibody with alkaline phosphatase, and detecting a diseased plant to be detected by a DAS-ELISA (double antibody sandwich-enzyme-linked immunosorbent assay) method. The method for detection of TYLCV in the invention has high sensitivity and can be used for qualitative and quantitative analysis, with a lowest detection limit of 9.75ng / mL; it also has simple and rapid operation that can be finished within several hours; with strong specificity and good repeatability, the method has very obvious immunological reaction to TYLCV; characterized by high reliability, the detection accuracy of the method is very close to PCR (polymerase chain reaction) method; the detection cost is low, and the reagent has small consumption amount and can be stored for a long time. Thus, the method provided in the invention is very suitable for large scale routine detection.

The invention discloses a Coxsackie virus A6 strain (WF057R) and applications thereof. The preservation number of the Coxsackie virus A6 strain (WF057R) in China General Microbiological Culture Collection Center (CGMCC) is CGMCC No.13393. A vaccine and CVA6 antiserum prepared from WF057R can treat and prevent diseases caused by CVA6. Furthermore, the maternal-transferred antibody of WF057R also has a protective effect on newborn mice. WF057R can also be used to establish a stable Coxsackie virus A6 infected animal model with good repeatability, and the animal model can be applied to drug antivirus treatment and immune protective effect evaluation of viral inactivation vaccines.

Genes encoding herpes simplex virus type 2 (HSV-2) proteins were cloned into eukaryotic expression vectors to express the encoded proteins in mammalian muscle cells in vivo. Animals were immunized by injection of these DNA constructs, termed polynucleotide vaccines or PNV, into their muscles. In a DNA titration, it was found that a single immunization of ≧0.5 μg of (one) PNV, gave >90% seroconversion by ten weeks post immunization. Immune antisera neutralized both HSV-2 and HSV-1 in cell culture. When animals were challenged with HSV-2, significant (p<0.001) protection from lethal infection was achieved following PNV vaccination. DNA constructs may be full-length, truncated and / or mutated forms and may be delivered along or in combination in order to optimize immunization and protection from HSV infection.

Provided herein are a hybridoma cell line producing monoclonal antibody against foot-and-mouth disease virus (FMDV), the monoclonal antibody therefrom, reagent and kit for ELISA, and immunoassay method. The hybridoma cell line is produced by cell fusion of a parental cell and a myeloma cell line and has the same characteristics as the cell line whose strain designation is CmA40 and deposition number is ATCC (To be Provided). The parental cell is a splenocyte isolated from the spleen of a mouse immunized by an antigen derived from a 3ABC non-structural protein (NSP) of FMDV. The antigen used here is expressed by a prokaryotic cell. The monoclonal antibody produced by the hybridoma cell line can specifically recognize a 3ABC polypeptide and does not cross-react with an antiserum of swine vesicular disease virus.

The invention provides a kit for detecting serum amyloid A content and a preparation method and detection method thereof and belongs to the field of medical detection. The kit comprises a hemolytic agent R1 and a latex reagent R2 which are independent with each other. The latex reagent R2 comprises latex coated with a serum amyloid A antibody. The latex coated with the serum amyloid A antibody isprepared from latex having particle sizes of 60-130nm and coated with the anti-serum amyloid A1 antibody and latex having particle sizes of 190-220nm and coated with the anti-serum amyloid A2 antibodythrough mixing. The invention also provides a preparation method and a detection method of the kit for detecting serum amyloid A content. The kit can detect whole blood, serum, a cerebrospinal fluidand a pleuroperitoneal fluid, utilizes a small amount of samples, is free of separation or pretreatment, can be directly used for detection and has good sample compatibility.

The invention discloses a preparation method of immunogold rapid test paper for vibrio parahaemolyticus. The method comprises the following steps of: (a) preparing a vibrio parahaemolyticus antigen; (b) preparing a vibrio parahaemolyticus inactivated vaccine; (c) preparing a polyvalent antiserum with a vaccine immune experimental rabbit; (d) purifying the antiserum; (e) marking with colloidal gold; and (f) preparing an immunogold test strip. A detection technology in which the immunogold rapid test paper for vibrio parahaemolyticus is adopted has the advantages of easiness, sensitivity, rapidness, specificity and the like; the customs clearance speed of port imported and exported goods can be increased greatly; and meanwhile, the test paper has the advantages of low price, no need of instrument, easiness and rapidness for operationing and easiness in judging results.

This invention discloses monoclonal antibody against anti-LCDV immunoglobulin of Paralichthys olivaceus, which is excreted by hybridoma JF-lgM-H (CCTCC-C200631). The method comprises: immunizing Paralichthys olivaceus with LCDV inactivated by formalin to prepare antiserum, purifying Paralichthys olivaceus immunoglobulin, immunizing Balb / c mice as antigen, preparing hybridoma cells by cell engineering method, and screening the monoclonal antibody by immunoassay. Indirect ELISA and indirect immunofluorescent antibody assay show that this monoclonal antibody is located on the heavy chain (7-80 kDa) of the anti-LCDV immunoglobulin. The monoclonal antibody can be used for preparing reagents for detecting LCDV infection in early stage, and evaluating the immune effects of LCDV vaccine inactivated by formalin.