22 results about "Marker Antigens" patented technology

This application relates to a detection reagent, kit, and method that simultaneously improves sensitivity and broadens the detection range in competitive immunoassay. The reagent includes a detection antibody bound to luminescent microspheres that specifically binds to analyte molecules in the sample, and two sets of labeled antigens containing the same competing antigen and labeled molecules of different molecular weights bound to it. The competing antigen competes with the analyte molecule for binding to the detection antibody, and the affinity of the competing antigen for binding to the detection antibody is higher than that of the analyte molecule for binding to the detection antibody. The technical solution of this application, by using two sets of labeled antigens with different affinities, ensures that the labeled antigens that play a dominant role in samples with varying concentrations of analyte molecules differ. This is suitable for competitive immunoassay of small molecule antigens or haptens in both low-concentration and high-concentration samples, and can broaden the detection range while ensuring high functional sensitivity.

The invention relates to the field of CAR-T cell concentration detection, in particular to a method for detecting CAR-T cells through combination of microfluidics and immunofluorescence. Through immunomagnetic bead enrichment and micro-fluidic chip partitioning, CAR-T cells are separated from a blood sample through immunomagnetic beads, then labeling is carried out or enhanced by adding a labeling antigen, anti-TCR-PE and DAPI, three-channel fluorescence labeling is achieved, quantitative analysis can be carried out on the number, the cell morphology and the function of the CAR-T cells through automatic immunofluorescence imaging, and the accuracy of CAR-T cell detection is improved. And the detection limit reaches 0.001%.

The present invention proposes a microfluidic chip for the combined detection of C-reactive protein and ferritin and its application. The chip comprises a substrate and a cover sheet pressed onto the substrate. The substrate and cover sheet enclose a microchannel. The left end of the microchannel is connected to a buffer injection hole provided on the cover sheet, and the right end is connected to a waste liquid area. The waste liquid area is provided with a removable absorbent paper sheet. The microchannel is provided with a labeling area, a detection area, and a sample injection area from left to right. The labeling area is provided with a labeled anti-human CRP antibody, a labeled anti-human SF antibody, a labeled second antibody, and a labeled antigen. The first detection site is provided with an anti-human CRP antibody, the first reference point is provided with a first antibody that reacts with the labeled second antibody, the second detection site is provided with an anti-human SF antibody, and the second reference point is provided with an antibody that reacts with the labeled antigen. This can simplify the detection process of C-reactive protein and ferritin and shorten the detection time.

The invention discloses a chemiluminiscence detection reagent for detecting an HCV antibody and a use method of the chemiluminiscence detection reagent, and relates to the technical field of immunodetection. The detection reagent is combined with a sandwich method and an indirect method for detection. When in use, a sample is incubated with the first reagent and the sample treating fluid, so that the HCV antibody to be detected in the sample reacts with the HCV coating antigen in the first reagent to form an antigen-antibody compound; adding a second reagent, and reacting the formed antigen-antibody compound with the HCV labeled antigen labeled by acridinium ester and the anti-IgG antibody labeled by acridinium ester in the second reagent to form an antigen-antibody-antigen compound and an antigen-antibody-secondary antibody compound; and finally, adding excitation liquid to enable acridinium ester to emit light, and detecting the HCV antibody in the sample by detecting a light-emitting signal value. The technical scheme of the invention greatly improves the sensitivity and specificity of HCV antibody detection, and can be used for solving the problem of easy missing detection in HCV antibody detection.

This invention proposes a microfluidic chip and its application based on dual-color and dual-layer technology to increase detection sites. The chip includes a substrate and a cover plate laminated onto the substrate. The substrate and cover plate enclose a microchannel. The microchannel is provided with a labeling region and a detection region. The labeling region includes a first labeling region on the lower surface of the cover plate and a second labeling region on the upper surface of the substrate. The detection region includes multiple detection sites on the lower surface of the cover plate and the upper surface of the substrate. The first labeling region is coated with a first labeled antigen / antibody, and the second labeling region is coated with a second labeled antigen / antibody. The first and second labeled antigens / antibodies are fluorescent microspheres labeled with two fluorophores that have the same excitation spectrum but different emission spectra. The multiple detection sites are respectively coated with capture antigens / antibodies that bind to different target analytes. By using a layered design combined with dual-color labeling, the number of simultaneously detectable indicators within the microchannel is increased.

The application provides an anti-LL37 antibody detection kit, which comprises a magnetic particle-carbamylated LL37 protein complex suspension, a mixed solution of biotin-PEGn-carbamylated LL37 protein and alkaline phosphatase coupled with streptavidin and a chemiluminescent substrate solution, wherein the sequence of the carbamylated LL37 protein is: {L-HomoCit}LGDFFR{K-HomoCit}S{K-HomoCit}E{K-HomoCit}IGKEF{K-HomoCit}RIVQRI{K-HomoCit}DFLRNLVP RTES. The application is based on a double antigen sandwich method, and the capture efficiency of the anti-LL37 antibody in serum is improved by designing the sequences of the coated antigen and the labeled antigen, so that the LL37 antibody in serum can be detected rapidly and efficiently.

The present invention relates to a method of detecting liver cancer in a mammalian subject by detecting an antibody in a test sample comprising a bodily fluid from the mammalian subject, wherein the antibody is an autoantibody immunologically specific for a tumour marker protein selected from the group consisting of MMP9, AIF1, EpCAM and CDKN1B, which method comprises contacting the test sample with a tumour marker antigen selected from the group consisting of MMP9, AIF1, EpCAM and CDKN1B and determining the presence or absence of complexes of the tumour marker antigen bound to autoantibodies present in the test sample where the presence of said complexes is indicative of the presence of liver cancer. Also included within the invention are corresponding methods of diagnosing and treating liver cancer in a mammalian subject, corresponding methods of predicting response to an anti-liver cancer treatment, a corresponding method of detecting an antibody in a test sample comprising a bodily fluid from a mammalian subject and kits suitable for performing methods of the invention.

The invention provides a micro-fluidic chip for increasing detection sites based on double-color and double-layer technology and application thereof, the micro-fluidic chip comprises a substrate and a cover plate pressed on the substrate, the substrate and the cover plate enclose to form a micro-channel, the micro-channel is provided with a marking area and a detection area, and the marking area and the detection area are arranged on the substrate. The marking area comprises a first marking area arranged on the lower surface of the cover plate and a second marking area arranged on the upper surface of the substrate, the detection area comprises a plurality of detection sites arranged on the lower surface of the cover plate and the upper surface of the substrate, the first marking area is coated with a first marking antigen / antibody, and the second marking area is coated with a second marking antigen / antibody. The first labeling area is coated with a first labeling antigen / antibody, the second labeling area is coated with a second labeling antigen / antibody, the first labeling antigen / antibody and the second labeling antigen / antibody are two fluorescent microsphere labels with the same excitation spectrum and different fluorescein emission spectrums, and the plurality of detection sites are respectively coated with capture antigens / antibodies combined with different target analytes. The number of synchronous detection indexes in the micro-channel is increased through layered design and combination of double-color marks.

The application discloses a fluorescence resonance energy transfer probe system and a method for high-sensitivity homogeneous immuno-detection of mycophenolic acid in whole blood. The application uses NaYbF4:Tm nanocrystals marked with BA monoclonal antibody on the surface as a donor probe, NaDyF4 nanocrystals marked with BA antigen on the surface as an acceptor probe, and constructs a fluorescence resonance energy transfer probe system. The probe system is used for qualitative and quantitative detection of mycophenolic acid in whole blood. The donor probe is incubated with a whole blood sample to be detected, and the fluorescence intensity emitted at 800 nm is tested. Then, the acceptor probe is added for joint incubation, and the fluorescence intensity emitted at 800 nm is tested. The ratio of the fluorescence intensity of the two times is compared with a control group or a standard curve, so that the mycophenolic acid in the whole blood can be qualitatively or quantitatively detected. The probe system can effectively overcome the background interference of the whole blood sample, and realize high-sensitivity homogeneous immuno-detection of BA in the whole blood.

The invention discloses a bigeminy immunoassay kit for detecting an HPV (human papillomavirus) antibody and a preparation method of the bigeminy immunoassay kit, relates to the field of biomedicine, and is characterized in that the bigeminy immunoassay kit for detecting the HPV antibody and the preparation method of the bigeminy immunoassay kit are provided. The magnetic microspheres for detecting the neutralizing antibodies of the human papilloma viruses 16 and 18 are coated with an HPV16 antibody and an HPV18 antigen, acridinium ester is used for marking the HPV16 antigen and the HPV18 antibody, an in-vitro diagnostic kit for detecting the neutralizing antibodies of the human papilloma viruses 16 and 18 is established on the basis, and a chemiluminescence immunoassay method is combined for analysis. The bigeminy immunoassay kit for quantitative detection of the HPV antibody is high in sensitivity, good in specificity, simple and convenient to operate and appropriate in price.

Disclosed herein are immunoassays for detecting an antibody in a biological sample from a subject and / or diagnosing an autoimmune disease in a subject. The disclosed immunoassays use a single, direct step to assess the level of antibody in a biological sample from the subject by simultaneously binding the antibody to a capture antigen (e.g. an unlabeled antigen bound to a solid support) and a labeled antigen not bound to the solid support.

The invention discloses an enzyme-labeled antigen for HIV diagnosis, the enzyme-labeled antigen is obtained by carrying out a directional coupling reaction on a gp41 recombinant antigen and an enzyme, the sequence of the gp41 recombinant antigen is shown as SEQ ID NO: 1, the enzyme is alkaline phosphatase, and the gp41 recombinant antigen and the enzyme are separated from each other. The preparation method of the enzyme-labeled antigen comprises the following steps: genetic engineering expression and purification of the antigen, modification of the antigen, activation of alkaline phosphatase, directional coupling reaction and stability optimization. The HIV recombinant antigen-alkaline phosphatase marker prepared by the invention is high in activity, strong in specificity, good in stability and good in inter-batch uniformity.

This invention proposes an immunomicrofluidic chip for the joint detection of IgM and IgG antibodies against the same pathogen and its application. The chip includes a substrate and a cover plate pressed onto the substrate. The substrate and cover plate together form a microchannel. From left to right, the microchannel has a labeled area and a detection area. The detection area includes an IgG detection area and an IgM detection area. The upper surface of the substrate, corresponding to the labeled area, is coated with labeled anti-human IgG antibody, and the upper surface of the substrate, corresponding to the IgG detection area, is coated with recombinant pathogen antigen. The lower surface of the cover plate, corresponding to the labeled area, is coated with labeled antigen, and the lower surface of the cover plate, corresponding to the IgM detection area, is coated with anti-human IgM antibody. By integrating the IgM antibody capture detection area and the IgG antibody indirect detection area into separate fluid layers of the chip, simultaneous but non-interfering detection of IgM and IgG antibodies in the same channel can be achieved after a single sample injection.

The invention relates to a fusion expression protein antigen combination containing a mycobacterium tuberculosis antigen TB16-38 and a fusion expression protein antigen combination containing a mycobacterium tuberculosis antigen and a mycobacterium bovis antigen MPB83-CFP10-ESAT-6. The method for preparing the combined antigen comprises the following steps: connecting a mycobacterium tuberculosis 38KD gene and a 16KD gene to escherichia coli by adopting a linker, expressing to obtain a TB16-38 antigen, and connecting a mycobacterium tuberculosis ESAT-6 gene, a CFP10 gene and a mycobacterium bovis MPB83 gene to escherichia coli by adopting two linkers, and expressing to obtain an MPB83-CFP10-ESAT-6 antigen; the invention relates to a mycobacterium tuberculosis and mycobacterium bovis antibody detection method, which comprises the following steps: fixing two labeled antigens on a solid phase carrier A together, contacting the carrier with a sample to be detected, moving the obtained labeled antigen-antibody compound on a solid phase carrier B, respectively contacting with two antigens (T1 and T2) independently coated on the carrier B, and fixing at the positions of T1 (6) and T2 (7), and determining mycobacterium tuberculosis and mycobacterium bovis antibodies according to display signals of T1 (6) and T2 (7).

The invention provides a method for preparing the same pathogen IgMamp; the immune micro-fluidic chip comprises a substrate and a cover plate pressed on the substrate, the substrate and the cover plate define a micro-channel, the micro-channel is sequentially provided with a marking area and a detection area from left to right, the detection area comprises an IgG detection area and an IgM detection area, the position, corresponding to the marking area, of the upper surface of the substrate is coated with a marked anti-human IgG antibody, and the IgM detection area is coated with an anti-human IgG antibody. The position, corresponding to the IgG detection area, of the upper surface of the substrate is coated with a pathogen recombinant antigen, the position, corresponding to the marking area, of the lower surface of the cover plate is coated with a marking antigen, and the position, corresponding to the IgM detection area, of the lower surface of the cover plate is coated with an anti-human IgM antibody. An IgM antibody capture detection area and an IgG antibody indirect detection area are respectively integrated on independent fluid layers of the chip, and synchronous but non-interfering detection of IgM and IgG antibodies in the same channel can be realized after a single sample is injected.

The invention provides a micro-fluidic chip for combined detection of hepatitis A / hepatitis E virus IgM antibody and application thereof, the micro-fluidic chip comprises a substrate and a cover plate pressed on the substrate, the substrate and the cover plate enclose to form a micro-channel, the micro-channel is provided with a marking area and a detection area, and the marking area and the detection area are arranged on the substrate. The marking area comprises a first marking area arranged on the lower surface of the cover plate and a second marking area arranged on the upper surface of the substrate, and the detection area comprises an HEV-IgM antibody detection area arranged on the lower surface of the cover plate and an HAV-IgM antibody detection area arranged on the upper surface of the substrate. An IgG antibody capture area is arranged on the upper surface of the substrate corresponding to the position right below the sample injection hole, the IgG antibody capture area is provided with a magnetic microsphere-anti-human IgG antibody, the first marking area is provided with a marked HEV antigen, the second marking area is provided with a marked anti-human IgM antibody, the HEV-IgM antibody detection area is provided with an anti-human IgM antibody, and the HAV-IgM antibody detection area is provided with an HAV antigen. A capture method and an indirect method are combined to realize joint detection of the IgM antibodies related to the two viruses.

The invention relates to a method for labeling antigen protein by biotin. The method comprises the following steps: (1) providing a ligase compound formed by Ni < 2 + > chelated polypeptide and recombinant Turbo ID protein; (2) providing a recombinant antigen protein with HisTag (HisTag); and (3) the recombinant antigen protein with HisTag reacts with biotin in the presence of the ligase complex, ATP and Mg < 2 + > to obtain the biotin-labeled antigen protein, in the step (1), the amino acid sequence of the polypeptide is shown as SEQ ID NO: 1, and the amino acid sequence of the recombinant Turbo ID protein is shown as SEQ ID NO: 2. According to the method disclosed by the invention, the labeling efficiency when the antigen protein is labeled by biotin is improved, the biotin utilization rate is improved, and the positive detection rate of specific detection of the antibody in a clinical sample is improved.

The invention discloses a fluorescence resonance energy transfer probe system and a method for high-sensitivity homogeneous immunodetection of rice yeast acid in whole blood. The method comprises the following steps: constructing a fluorescence resonance energy transfer probe system by taking a NaYbF4: Tm nanocrystal of which the surface is marked with a BA monoclonal antibody as a donor probe and taking a NaDyF4 nanocrystal of which the surface is marked with a BA antigen as a receptor probe; the probe system is used for qualitative and quantitative detection of rice yeast acid in whole blood, the donor probe and a whole blood sample to be detected are incubated, the intensity of fluorescence emitted at the 800 nm position is tested, then the receptor probe is added for co-incubation, and the intensity of fluorescence emitted at the 800 nm position is tested. The ratio of the two fluorescence intensities is compared with a control group or a standard curve, so that the rice yeast acid in the whole blood can be qualitatively or quantitatively detected. The probe system disclosed by the invention can effectively overcome background interference of a whole blood sample and realize high-sensitivity homogeneous immunodetection of BA in whole blood.

Proposed are a disease test kit for testing a disease on the basis of a body fluid and a disease testing method using the kit, wherein the disease test kit comprises: a sample pad to which a first blocking buffer is attached and into which a body fluid sample is introduced; a conjugate pad to which a second blocking buffer is attached and which binds the body fluid sample and a labeling antibody while storing the labeling antibody; a membrane pad for detecting a concentration of a labeling antigen in the body fluid sample bound to the labeling antibody; and a test strip including an absorption pad for absorbing and storing the sample that has passed through the membrane.

This invention proposes a microfluidic chip for the combined detection of hepatitis A / hepatitis E virus IgM antibodies and its application. The chip includes a substrate and a cover plate pressed onto the substrate. The substrate and cover plate enclose a microchannel, which is provided with a labeling area and a detection area. The labeling area includes a first labeling area on the lower surface of the cover plate and a second labeling area on the upper surface of the substrate. The detection area includes an HEV-IgM antibody detection area on the lower surface of the cover plate and a HAV-IgM antibody detection area on the upper surface of the substrate. An IgG antibody capture area is provided on the upper surface of the substrate directly below the sample injection well. The IgG antibody capture area contains magnetic microspheres containing anti-human IgG antibodies. The first labeling area contains labeled HEV antigen, the second labeling area contains labeled anti-human IgM antibody, the HEV-IgM antibody detection area contains anti-human IgM antibody, and the HAV-IgM antibody detection area contains HAV antigen. The capture method and the indirect method are used in parallel to achieve the combined detection of IgM antibodies related to the two viruses.

This invention relates to the field of biotechnology, specifically to a composite quality control product for gastric tumor markers and its preparation method. The composite quality control product comprises gastric tumor marker antigens and a quality control buffer solution. The quality control buffer solution comprises 1-10% protein protectant, 0.01-3% surfactant, 2-20% thickening antioxidant, 0.5-500 mg / L protease inhibitor, 0.01-3% metal ion chelating agent, and 0.5-10% buffer matrix. The gastric tumor marker antigens include G17, PGII, and CA72-4. This invention, by addressing the interactions between G17, PGII, and CA72-4 antigens, achieves composite quality control for eight gastric tumor markers. This covers the quality control of four conventional gastric tumor detection reagents (PGI, PGII, G17, and CA72-4) and four broad-spectrum tumor detection reagents (AFP, CEA, CA19-9, and CA242), which is of significant importance for clinical gastric tumor screening.

The application relates to a method for labeling an antigen protein with biotin, which comprises the following steps: (1) providing a ligase complex formed by a polypeptide chelated with Ni 2+ and a recombinant TurboID protein; (2) providing a recombinant antigen protein with a HisTag; and (3) reacting the recombinant antigen protein with the HisTag with biotin under the condition that the ligase complex, ATP and Mg 2+ exist to obtain the antigen protein labeled with biotin, wherein the amino acid sequence of the polypeptide in the step (1) is shown in SEQ ID NO: 1, and the amino acid sequence of the recombinant TurboID protein is shown in SEQ ID NO: 2. The method improves the labeling efficiency of the antigen protein labeled with biotin, improves the biotin utilization rate, and improves the positive detection rate of specific antibody detection in a clinical sample.