143 results about "Antibody labeling" patented technology

The invention provides a micro-fluidic chip for combined detection of C-reactive protein and ferritin and application thereof.The micro-fluidic chip comprises a substrate and a cover plate pressed on the substrate, a micro-channel is defined by the substrate and the cover plate, the left end of the micro-channel is communicated with a buffer solution sample adding hole formed in the cover plate, the right end of the micro-channel is communicated with a waste liquid area, and the left end of the micro-channel is communicated with a buffer solution sample adding hole formed in the cover plate. The waste liquid area is provided with a movable water-absorbing paper sheet, the microchannel is sequentially provided with a marking area, a detection area and a sample adding area from left to right, the marking area is provided with a marked anti-human CRP antibody, a marked anti-human SF antibody, a marked second antibody and a marked antigen, the first detection site is provided with an anti-human CRP antibody, and the second detection site is provided with an anti-human SF antibody. The first reference point is provided with a first antibody which 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 which reacts with the labeled antigen. The detection process of C-reactive protein and ferritin can be simplified, and the detection time is shortened.

The invention relates to an anti-VIM nano antibody and application thereof, belongs to the technical field of biology, and aims to solve the problem of Vimentin VIM detection, and has the key points that a complementary determining region CDR of the nano antibody comprises CDR1, CDR2 and CDR3 sequences: (I): (1) the amino acid sequence of the CDR1 is shown as SEQ ID NO.27, and (2) the amino acid sequence of the CDR2 is shown as SEQ ID NO.33; and (3) the amino acid sequence of the CDR3 is shown as SEQ ID NO.39. The nano antibody and the polypeptide thereof have the effects that the nano antibody and the polypeptide thereof have very high affinity and activity, can specifically recognize and combine with VIM, can be used for capturing and detecting VIM positive cells, and can be applied to immunofluorescence analysis or immunohistochemical analysis and the like through a proper antibody labeling technology.

The invention belongs to the technical field of gene engineering, and particularly relates to a Cre reporting system based on single-domain antibody labeling and application of the Cre reporting system. According to the shark single-domain antibody with Cre specificity, when a DNA sequence of dLox-EGFP is provided, Cre can be marked and reported in real time, and reporter genes can be expressed only when Cre exists in cells, so that a strictly regulated gene expression platform is provided. The mark is completely reversible and is completely compatible with the currently used Cre / loxp system.

The invention relates to the technical field of pesticide detection, and discloses a multi-component colloidal gold test strip for rapid detection of organophosphorus as well as a preparation method and application thereof, the preparation method comprises the following steps: preparing a specific antibody of the organophosphorus, and preparing a colloidal gold solution at the same time; mixing and incubating the specific antibody and a colloidal gold solution to obtain an antibody-labeled colloidal gold solution; pre-treating a detection pad, and immersing the detection pad in the antibody-labeled colloidal gold solution to obtain an impregnated pad; and sequentially assembling the dried dipping pad, a water absorption pad, a sample pad and a combination pad on a bottom plate to form a complete test strip. By improving the preparation process of the antibody and the assembly technology of the test strip, the detection sensitivity and specificity are remarkably improved. The test strip is simple and convenient in preparation process, high in cost benefit and convenient for large-scale production. In addition, the test strip is convenient to use and does not need to depend on professional equipment or technicians, and the detection result is visual and convenient to read.

The invention provides a kit and a method for simultaneously detecting CEA, CA125 and CA153 based on a flow cytometry, which are characterized in that proper microspheres and antibody markers are specially arranged for CEA, CA125 and CA153 antibodies, a 1.5-step incubation method and a biotin system are combined, and proper blocking agents and confining liquid are added into a reaction buffer solution, so that the CA153 in a sample is better combined with the microspheres and the antibody markers, and the detection accuracy is improved. According to the present invention, the cross influence of the CA153 on the CEA and the CA125 is reduced, such that the detection accuracy can be significantly improved when the three tumor markers such as the CEA, the CA125 and the CA153 are simultaneously detected based on the flow cytometry, the detection sensitivity of the CEA can achieve 0.3 ng / mL, the detection sensitivity of the CA125 can achieve 0.5 U / mL, and the detection sensitivity of the CA153 can achieve 0.5 U / mL so as to meet the clinical requirements.

The invention discloses a preparation method of a triple detection kit for influenza A, influenza B and new coronavirus antigens, and belongs to the field of respiratory virus in-vitro detection. In the antibody labeling stage, the antigen and the antibody can be more sufficiently combined in a manner of adding the linking arm, so that the antigen detection sensitivity can be improved. In the metal spraying stage, a novel coronavirus monoclonal antibody, an influenza A virus monoclonal antibody and an influenza B virus monoclonal antibody marked by latex microspheres are sequentially subjected to multi-layer metal spraying at the same position by a metal spraying instrument, the two antibodies are isolated by a trehalose solution, and metal spraying of three layers of antibodies isolated from each other is completed. And the cross reaction between different antibodies of the conjugate pad in the storage process is avoided, so that non-specific binding is avoided, and the storage stability is improved. The detection kit prepared by the method can specifically detect the novel coronavirus, the influenza A virus and the influenza B virus, and has the advantages of high sensitivity, good repeatability and the like.

The invention discloses a serum amyloid protein A immunochromatography detection reagent strip which comprises a reaction bottom plate, a sample pad, a combination pad, a chromatography membrane and absorbent paper, the chromatography membrane is arranged in the middle of the reaction bottom plate, the sample pad and the absorbent paper are arranged at the two ends of the reaction bottom plate respectively, the chromatography membrane is arranged in the middle of the reaction bottom plate, the combination pad is arranged between the chromatography membrane and the sample pad, the sample pad is partially overlapped on the combination pad, and the combination pad is partially overlapped on one side of the chromatography membrane. The absorbent paper is partially overlapped on the other side of the chromatography membrane; quantum dot fluorescent microspheres used for serum amyloid protein A antibody labeling and quantum dot fluorescent microspheres used for DNP-BSA labeling are solid-phase on the combination pad; the chromatographic membrane is provided with a detection line coated with a serum amyloid protein A detection antibody and a quality control line coated with a rabbit anti-DNP quality control antibody; and a detection line and a quality control line are sequentially arranged on the chromatography membrane in parallel at intervals along the chromatography direction. The invention further discloses a corresponding preparation method, a detection kit, a detection method and application.

This invention belongs to the field of microfluidics, and particularly relates to the use of a pine pollen microrobot combined with a microfluidic chip for cell capture. The pine pollen microrobot is gold-sputtered pine pollen, and the microfluidic chip is injected with the pine pollen microrobot and a conductive cell solution. When an alternating current signal is applied to the microfluidic chip, the fluid surrounding the pine pollen microrobot generates an induced charge electroosmotic vortex, causing cells in the cell solution to move toward the cavity of the pine pollen microrobot, thereby achieving cell capture. Using induced charge electroosmosis (ICEO) technology, this invention enables the efficient capture of blood cells and other micron-sized cell structures. This solves the problem of traditional capture techniques relying on antibody labeling and mechanical capture, reduces operational complexity and capture difficulty, and improves capture success rate and raw material utilization.

The invention discloses a culture method for inducing high expression of umbilical cord mesenchymal stem cells (LRP1) and application, and belongs to the technical field of biological medicines. The technical problem to be solved is to provide a standardized culture method for directionally inducing a specific functional phenotype cell population, so that efficient and accurate induction of LRP1 high-expression UCMSCs is realized, and clinical adaptation is met. According to the key point of the technical scheme, the culture method for inducing the LRP1 high-expression umbilical cord mesenchymal stem cells comprises the following steps: inoculating umbilical cord mesenchymal stem cells into a culture dish at the inoculation density of 5.5 * 10 < 3 > / cm < 2 > to 3.6 * 10 < 4 > / cm < 2 >, and culturing for 24-72 hours by using a special mesenchymal stem cell culture medium without adding a supplement, so that the acquisition efficiency of the LRP1 high-expression UCMSCs is improved, and the yield of the LRP1 high-expression UCMSCs is increased. Antibody labeling and flow sorting are not needed, so that the operation steps are simplified, and the complexity and cost of the culture process are reduced.

This invention discloses a quantitative test strip for HBV PreS1 antibody based on the AIE molecule, its preparation method, and its applications, relating to the fields of molecular biology, immunology, and biodetection technology. The test strip includes a sample pad, a labeling pad, a nitrocellulose membrane (NC membrane), an absorbent pad, and a base plate. The NC membrane detection line is coated with HBV PreS1 antigen to capture PreS1 antibodies in the sample. The labeling pad contains an immobilized Anti-Human IgG antibody conjugated to the AIE molecule, which binds to the PreS1 antibodies in the sample to form an immune complex. The test strip of this invention enables rapid detection of PreS1 antibodies and achieves enhanced fluorescence in the detection results. Combined with a fluorescence reader or fluorescence card reader, the detection results can be digitally and quantitatively analyzed. It is simple to operate, rapid in detection, and significantly improves detection sensitivity and enables quantitative analysis.

The invention belongs to the technical field of immunochemiluminescence, and relates to a PAMAM-mediated antibody marker and a synthesis method and application thereof, the PAMAM-mediated antibody marker is composed of an antibody, PAMAM and a marker, amino groups on the surface of the antibody can be converted into sulfydryl groups, and the marker is provided with succinimide groups which can react with the amino groups. The steric hindrance of antigen-antibody reaction is reduced through PAMAM mediation, the labeling proportion is increased to the maximum extent, and a high coupling effect can be achieved with a small feeding proportion.

The invention relates to a method for detecting activation of specific T cells after tuberculosis infection and application of the method. Specifically, the invention provides a reagent combination for detection, and the reagent combination comprises an anti-CD4 antibody (CD4 antibody), an anti-CD154 antibody (CD154 antibody), an IFN-gamma detection antibody and a tuberculosis specific antigen. According to the method, through peripheral whole blood collection, PBMC cell separation, T cell activation (activation), activation termination (blocking), antibody labeling, membrane rupture, fixation, intracellular cytokine dyeing, flow cytometry detection and result analysis and judgment, the ratio of the expression IFN-gamma cells in the CD4 + CD154 + cells can be rapidly and accurately determined, and by combining an ROC curve and a constructed judgment standard, the content of the IFN-gamma cells in the CD4 + CD154 + cells can be determined, and the content of the IFN-gamma cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells can be determined. And the sample to be detected is judged to be in a positive / negative / gray region, so that whether a patient is infected by the mycobacterium tuberculosis or not can be more accurately diagnosed, especially aiming at a whole blood sample from a special crowd.

The application discloses an acridinium ester labeled complex, which comprises a body, and a labeling group labeled on the body, wherein the body is an antibody, and the labeling group is an acridinium ester group. A preparation method of the acridinium ester labeled complex comprises the following steps: sucking a labeling buffer into a centrifugal tube; sucking an antibody to be labeled into the centrifugal tube and mixing to generate a first mixed solution; adding an acridinium ester solution and mixing, and then performing reaction at 25 DEG C in the dark for 2 hours to generate a second reaction solution; adding a termination buffer and mixing, and then performing reaction at 25 DEG C in the dark for 30 minutes to generate a third reaction solution; performing desalting purification on the third reaction solution to generate a fourth mixed solution; and filtering the fourth mixed solution with a 0.22 mu m filter membrane to generate the acridinium ester labeled complex. According to the preparation method, the labeling efficiency of the acridinium ester and the antibody is higher, the reactivity is stronger, the non-specific adsorption is lower, the sensitivity is higher, the repeatability and stability are better, and the labeling method is simple, rapid and easy to repeat.

This disclosure provides a method for analyzing peripheral blood leukocytes. In some embodiments, method may comprise: (a) labeling peripheral blood leukocytes isolated from a patient that has or is suspected of having inflammatory bowel disease (IBD) with a panel of distinguishably-labeled antibodies and (b) analyzing binding of the antibodies to the peripheral blood leukocytes. Kits for performing the method are also provided. Results from the analysis can be used to make a diagnosis of Crohn's disease or ulcerative colitis.

VIM-specific nanobody and its applications belong to the field of biotechnology. To solve the problem of detecting vimentin (VIM), the key point is that the complementarity-determining regions (CDRs) of the nanobody include CDR1, CDR2, and CDR3 sequences: (I): (1) The amino acid sequence of CDR1 is shown in SEQ ID NO.27; (2) The amino acid sequence of CDR2 is shown in SEQ ID NO.33; and (3) The amino acid sequence of CDR3 is shown in SEQ ID NO.39. The effect is that the nanobody and its polypeptide of the present invention have high affinity and activity, can specifically recognize and bind to VIM, can be used for the capture and detection of VIM-positive cells, and can be applied to immunofluorescence analysis or immunohistochemical analysis through appropriate antibody labeling techniques.

The invention discloses a kit and a method for detecting a biomarker p-Tau181 protein of Alzheimer's disease, and belongs to the technical field of in vitro detection.The kit comprises a magnetic bead working solution composed of magnetic beads labeled by a p-Tau181 antibody and a buffer solution, the visual non-magnetic microsphere working solution is composed of visual non-magnetic microspheres labeled by total tau antibodies and a buffer solution, the size of magnetic beads is 1-10 [mu] m, and the size of the visual non-magnetic microspheres is 1-10 [mu] m. The scheme has the advantages of high sensitivity, good specificity and the like, the detection limit can be reduced to 0.01 pg / mL, and a non-invasive and reliable detection means is provided for the Alzheimer's disease.

The invention relates to the technical field of biological detection, and discloses a tetrodotoxin colloidal gold detection card, which comprises a detection box used for packaging a detection assembly, the display groove is formed in the detection box and is used for observing a detection result; the liquid dropping groove is positioned at one end of the detection box and is used for accommodating a to-be-detected sample; and the detection paper is arranged in the detection box, comprises a nitrocellulose membrane, an adsorption pad and a diffusion pad, is provided with a detection area and a control area, and is used for displaying a detection result of tetrodotoxin. By optimizing the size of the gold nanoparticles and an antibody labeling process, the sensitivity and the specificity of the detection card are remarkably improved, and the low-concentration tetrodotoxin can be rapidly and accurately detected. Due to the reasonable particle size and the application of the stabilizer, non-specific adsorption is reduced, the false positive rate is reduced, the reliability and consistency of detection results are ensured, clear detection results can be obtained in a shorter time, and the accuracy of rapid judgment in field application is ensured.

The invention relates to the technical field of clinical medical examination, in particular to an immunochromatography test strip for enhancing near-infrared two-region fluorescence through supramolecular host-guest interaction and a preparation method of the immunochromatography test strip. The invention provides a preparation method of an immunochromatographic test strip. The preparation method comprises the following steps: preparing a rare earth nanoparticle (RENPs) labeling solution; preparing a microsphere combination pad; preparing a sample pad; coating an antibody with a nitrocellulose membrane; and assembling and manufacturing the card. The aggregation phenomenon and partial fluorescence quenching effect caused in the surface modification process of the rare earth microspheres are reduced by utilizing an efficient surface modification strategy of a supramolecular host compound, the fluorescence luminescence property of the rare earth microspheres is effectively improved, and meanwhile, the antibody labeling efficiency of the rare earth microspheres is further improved by utilizing a supramolecular self-assembly strategy; the prepared test strip is obviously improved in analysis sensitivity, has the advantages of simplicity in operation, low cost and short detection time, and is suitable for rapid and sensitive detection of low-concentration markers in complex biological samples.

The invention relates to a method for amplifying an acridine system of a chemiluminescence platform, and particularly provides a method for enhancing the luminous intensity in chemiluminescence immunoassay, which comprises the step of enhancing the luminous intensity by using a composition, and the composition comprises biotin labeled by an anti-acridine analogue antibody and streptavidin labeled by an acridine analogue. According to the method, the sensitivity and the luminescence value are amplified by utilizing the specific binding of the biotin and the avidin and then utilizing the superposition of the specific affinity binding mode of the acridine analogue and the acridine analogue antibody, the sensitivity can be improved by 5-10 times on the existing acridine analogue platform, and the application prospect is excellent.

The invention discloses a universal test strip for rapid detection of multiple nucleic acid amplification products as well as a preparation method and application of the universal test strip, and belongs to the technical field of rapid detection of nucleic acid. The universal test strip comprises a reaction bottom plate, a sample pad, a combination pad, a chromatography membrane and a water absorption pad, wherein the combination pad is loaded with a marked first target antibody, a marked first quality control antibody or antigen; the chromatography membrane is provided with a detection line T coated with a second target antibody and a quality control line C coated with a second quality control antibody or antigen; the detection line and the quality control line are sequentially arranged on the chromatography membrane in parallel at intervals along the chromatography direction. The universal test strip has the remarkable advantages of simplicity and convenience in operation, high detection speed, high sensitivity, strong specificity, low cost and the like, is wide in application technical range, is suitable for various nucleic acid detection technical methods, has strong tolerance to sample inhibitors, and is particularly suitable for being combined with isothermal amplification rapid detection technologies such as RPA (recombinase polymerase amplification), LAMP (loop-mediated isothermal amplification) and the like.

The invention provides an interleukin 33 detection kit as well as a preparation method and application thereof. The interleukin 33 detection kit comprises a first antibody coupled to a magnetic fluorescent light-emitting microsphere, a second antibody labeled with biotin and fluorescein-labeled streptavidin, the first antibody strain and the second antibody strain are monoclonal antibodies for resisting interleukin 33, and interleukin 33 specific binding sites recognized by the first antibody strain and the second antibody strain are different. According to the kit disclosed by the invention, the interleukin 33 antigen is specifically combined with the magnetic fluorescent luminous microsphere formed by coupling the interleukin 33 capture antibody, then the magnetic fluorescent luminous microsphere is specifically combined with the biotin-labeled interleukin 33 detection antibody, the conjugate is combined with fluorescein-labeled streptavidin, fluorescence is emitted, and the content of the interleukin 33 in a sample to be detected is determined. The method is high in sensitivity, good in repeatability, short in reaction time, simple to operate and high in anti-interference performance.

The invention provides an indirect ELISA (Enzyme-Linked Immunosorbent Assay) antibody detection method and an indirect ELISA antibody detection kit based on pyruvate Kinase (PK), and relates to the technical field of parasite detection. The kit comprises a coating antigen Italiella mannici PK protein, a primary antibody, a labeled secondary antibody and an initiator solution, wherein the primary antibody is a mouse or cat anti-Italiella mannici PK protein antibody to be detected; the labeled secondary antibody can be combined with the primary antibody combined with the PK protein of the andrometrium mansoni; and the labeled second antibody is provided with a detection marker. The method comprises the following steps: cloning a PK protein gene into a prokaryotic expression vector PQE80L (+), constructing a recombinant plasmid PQE80L-PK, inducing PK recombinant protein expression by utilizing IPTG (isopropyl-beta-d-thiogalactoside), purifying the recombinant protein to coat an elisa plate, and establishing an indirect ELISA (enzyme-linked immuno sorbent assay) detection method of the Mangnocorynth PK protein antibody by optimizing reaction conditions. The detection method has the advantages of high sensitivity and strong specificity, and provides support for the development of the detection technology of the sitonia mansoni and the prevention and control of the schizocyst.

The present invention discloses a 25-OH-D, P1NP and β-CTX joint detection kit, including a reagent card and a dissociating agent; the reagent card includes a PVC rubber plate, a nitrocellulose membrane arranged on the PVC rubber plate, a conjugation pad, a sample pad, and a blotting paper sequentially arranged on the nitrocellulose membrane; the nitrocellulose membrane is coated with 25-OH-D antibody, P1NP antibody, β-CTX antibody and quality control antibody; the conjugation pad is marked with 25-OH-D antibody, P1NP antibody, and β-CTX antibody tracer by a coupling method; the dissociating agent is used to dissociate 25-OH-D in the form of a binding protein in serum or plasma, and does not affect the detection results of P1NP and β-CTX. A preparation method of the kit is also disclosed. The present invention can perform joint quantitative detection of 25-OH-D, P1NP, and β-CTX in blood.

This invention relates to the field of biological detection, and more particularly to the preparation method and application of the t-PAI·C detection kit. The invention provides a detection reagent comprising: antibody-coated magnetic microparticles and enzyme-labeled antibodies; the antibodies include: tissue plasminogen activator-plasminogen activator inhibitor-1 complex antibody. In the detection reagent provided by this invention, when the sample contains the tissue plasminogen activator-plasminogen activator inhibitor-1 complex, the antibody coated with the magnetic microparticles and the enzyme-labeled antibody undergo an immune reaction with the complex, forming a sandwich-like immune complex consisting of the antibody coated with the magnetic microparticles, the complex, and the enzyme-labeled antibody; the enzyme on this complex acts on a luminescent substrate and emits light, and the concentration of the complex in the sample is calculated by detecting the luminescence signal.

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 provides an individualized drug screening method for leukemia based on CD45 monoclonal antibody enrichment. The method comprises the following steps: collecting a bone marrow extract sample or a peripheral blood sample, separating to obtain a mononuclear cell, labeling the mononuclear cell from a patient by adopting a human CD45 antibody, enriching a leukemia cell population through the CD45 monoclonal antibody based on a CD45 / SSC gating method, adding a leukemia primary drug sensitivity detection culture medium, and detecting the cell activity inhibition condition after adding a corresponding drug to act. The leukemia cell population is enriched through the CD45 monoclonal antibody based on CD45 / SSC, the cells are subjected to drug sensitivity detection to screen out leukemia individualized drugs, interference of normal cells in different quantity proportions in a source sample on leukemia drug sensitivity is eliminated, and response of the source leukemia cells to the drugs can be reflected more truly and visually. The rapid and accurate leukemia individualized drug screening method has important significance for improving the treatment outcome of acute myelogenous leukemia.

The present invention provides a kit and application for joint detection of CA724 and uric acid, the kit includes a test card and a sample detection solution, the test card includes a test strip, the test strip includes a base plate, a sample pad, a conjugation pad, a nitrocellulose membrane and a water-absorbing pad, the sample pad, the conjugation pad, the nitrocellulose membrane and the water-absorbing pad are sequentially adhered to the base plate, the conjugation pad is adsorbed with uric acid antibody-labeled fluorescent microspheres and CA724 antibody-labeled fluorescent microspheres, the nitrocellulose membrane is sequentially provided with a T1 detection line coated with uric acid-bovine serum albumin hapten, a T2 detection line coated with CA724 monoclonal antibody and a quality control line C according to the sample flow direction, and the sample detection solution is composed of specific components. The kit of the present invention can realize simultaneous quantitative detection of CA724 and uric acid, and the result is accurate, the sensitivity is high, the operation steps are simplified, the sample dosage is reduced, the time is saved, the cost is saved, and it is conducive to early gout diagnosis and treatment.

The present invention discloses an immunoassay reagent based on AIE magnetically encoded microspheres and AIE dyes, and a preparation method and application thereof; the immunoassay reagent comprises an AIE magnetically encoded microsphere labeled with an antibody-1 and a water-soluble AIE dye labeled with an antibody-2. The present invention adds the AIE magnetically encoded microspheres labeled with an antibody-1 and the water-soluble AIE dye labeled with an antibody-2 to a test solution containing a corresponding antigen, and after the reaction, the AIE magnetically encoded microspheres are separated by magnetic attraction, and the encoding peak and fluorescence quantitative detection concentration are obtained by flow cytometry. The present invention utilizes the high-regionality of the AIE magnetically encoded microspheres and the signal amplification effect of the quantitative detection of the AIE molecules to realize long linear range detection in complex samples of multiple markers, and provides a new detection strategy for multiple marker immunoassay.

The invention relates to a synthesis method of an antibody-drug conjugate labeled by a position-selective antibody based on proximity effect. According to the present application, it is possible to provide a method for synthesizing an antibody-drug conjugate by introducing a drug position-selectively into an antibody using a pyridine oxime derivative (labeled mediating protein) introduced into a binding protein comprising a non-standard amino acid.

The invention discloses a urine-based kit for semi-quantitatively detecting the content of vitamin D and a detection method, which can be used for detecting a urine sample. The kit for semi-quantitatively detecting the content of vitamin D based on urine is obtained by combining a nitrocellulose membrane, an antibody labeling technology, an antibody-antigen specific binding reaction and an immunomembrane chromatography technology. The kit comprises a vitamin D detection reagent strip comprising sample filtering paper, a colloidal gold combination pad, a nitrocellulose membrane and absorbent paper which are arranged in sequence, has the characteristics of simplicity and convenience in operation, rapidness in detection, high sensitivity and strong specificity, and can be used for semi-quantitative detection and judgment of the concentration of vitamin D in urine; therefore, whether the content of the vitamin D in the urine is normal or not is accurately distinguished, efficient and sensitive vitamin D detection is achieved, and a new and reliable solution is provided for vitamin D detection.