31 results about "Fluoreszein" patented technology

Aspects described herein pertain to engineered chimeric antigen receptors (CARs) and compositions thereof having specificity and affinity for fluorescein containing ligands presented on the surface of tumor cells. Also provided herein are compositions including CARs further comprising a spacer arm and methods of making and using these compositions.

The invention discloses a kit and method for detecting leukemia and lymphoma based on full-spectrum flow cytometry, the kit comprises 25 antibodies, the antibodies are specifically bound with fluorescein respectively, and leukemia and lymphoma are detected through full-spectrum flow cytometry; the 25 kinds of antibodies comprise HLA-DR (human leukocyte antigen-DR), CD38, CD7, CD34, Lambda, CD19, CD64, CD14, CD5, CD123, CD16, CD20, Kappa, CD117, CD13, CD45, CD11b, CD2, CD10, CD8, CD15, CD4, CD3, CD56 and CD33. The kit comprehensively covers development stages of various lines of bone marrow cells, and common abnormal expressions of various leukemia, myelodysplastic syndromes and lymphoma, and can preliminarily screen various leukemia and lymphoma.

The invention provides fluorescein and a purification and synthesis method thereof, and relates to the field of compound preparation. The method comprises the following steps: carrying out first mixing and first reaction on 4-(aminomethyl) benzoic acid, triethylamine, a first organic solvent and trifluoroacetate to obtain a first intermediate; performing second mixing on the first intermediate, a second organic solvent, triethylamine and 2-succinimido-1, 1, 3, 3-tetramethylurea tetrafluoroborate, and performing second reaction to obtain a second intermediate; and carrying out third mixing on the second intermediate, amino FAM, a third organic solvent and triethylamine, and carrying out third reaction to obtain fluorescein. The synthesis and purification method is efficient, and fluorescein can be rapidly obtained without column chromatography; chemical raw materials used in the synthesis method are simple and easy to obtain, the cost is low, reaction conditions are mild, amplified production is easy, the purification method is simple and convenient to operate, and the obtained intermediate is high in stability.

The invention provides a method for rapidly detecting a neutrophil extracellular trapping net. The method comprises the following steps: collecting peripheral blood; the method comprises the following steps: taking peripheral blood, adding different fluorescein-labeled anti-CD66b and CtiH3 antibodies and a dsDNA dye SYTOX Green, uniformly mixing, and incubating; adding a red blood cell lysis buffer, uniformly mixing, and incubating at room temperature; adding PBS, uniformly mixing, and centrifuging; the method comprises the following steps of: removing supernatant, adding PBS (Phosphate Buffer Solution) to resuspend cells, detecting by a flow cytometer, detecting CD66b positive cells as neutrophils by adopting an APC channel, detecting CtiH3 as CtiH3 positive neutrophils by adopting a PE channel in combination with a neutrophil population, detecting dsDNA as dsDNA positive neutrophils by adopting an FITC channel in combination with the neutrophil population, and calculating a detection result according to an NETs formation index calculation formula. The invention also provides a system for evaluating the NETs of the sample to be tested. According to the invention, the detection time of NETs detection is shortened, and the stability and diagnosis efficiency of NETs detection are improved.

This invention discloses a detection kit and identification method for CTCs based on SCLC molecular typing using microfluidic chips and multiplex immunofluorescence probe technology. The kit includes a microfluidic chip, a biotin-labeled trapping agent, a balanced salt solution, a tissue cell fixation solution, a specific hybridization blocking solution, an active nuclear staining solution, and a diluent. It also includes a detection agent composed of key subtype markers of SCLC labeled with different fluorescein groups, SCLC-related marker detection antibodies, and leukocyte auxiliary identification markers, or a detection agent composed of key subtype markers of SCLC labeled with different fluorescein groups and leukocyte auxiliary identification markers. This application enables non-invasive, early, and sensitive detection and analysis of the expression of four immunomarkers involved in CTCs in the peripheral blood of patients: ASCL1, NEUROD1, POU2F3, and YAP1. It is suitable for early tumor screening and can also be used for pan-cancer immunotherapy guidance, immunotherapy efficacy monitoring, and prognostic assessment.

A population of universal natural killer (NK) cells derived from human pluripotent stem cells (hPSCs) and engineered to overexpress the transcription factor ID2, NFIL3, and / or SPI1 and, optionally, an anti-programmed death ligand 1 (PD-L1) chimeric antigen receptor (CAR) and an anti-fluorescein isothiocyanate CAR are provided. Methods of treating cancer in a subject using the population of universal NK cells are also provided.

The invention discloses a method for identifying chronic granulocyte immune typing by using a flow cytometer. The method comprises the following steps: incubating and purifying a to-be-detected sample and a fluorescein-labeled antibody composition; the method comprises the following steps: carrying out flow cytometry detection on resuspended cells: firstly, circling a liquid flow stable interval by utilizing Time / CD45, FSC INT / PEAK and FSC INT / SSC INT scatter diagrams, and removing sticky bodies and cell debris; all leukocyte groups are circled through combination of CD45 and SSC, and mononuclear cells are circled in all leukocyte gates through CD45 / SSC gate setting; excluding CD 19 + B lymphocytes; cD7 + T and NK cells are excluded; cD2 + T and NK cells are eliminated; immature and mature granulocytes of CD 16 and CD24 < + > are excluded through CD 16 / CD24 gate setting; the method comprises the following steps of: screening CD14-CD16, removing basophilic granulocytes of CD14-CD16-and residual NK (Natural Killer) cells, dividing a cross gate to determine the proportion of CD 14 + CD16-cMo cells, CD 14 + CD16 + iMo cells and CD 16 + CD 14 ncMo cells, and determining the proportion of the CD14 + CD16-cMo cells, the CD14 + CD16 + iMo cells and the CD16 + CD14 ncMo cells; cD34 / SSC gate setting is carried out, and CD34 + original cells are circled to calculate the proportion.

The invention relates to a nano drug delivery system, the nano drug delivery system comprises a core, a shell and a membrane, the core comprises hollow microspheres, the shell comprises an iron ion-containing component and a copper ion-containing component, and the membrane comprises a cell membrane inserted with an artificial peptide; the shell is fixed on the surface of the core through ion adsorption, and the membrane wraps the surface of the shell; the hollow microspheres comprise polylactic acid-glycolic acid copolymer-ketal thiol-polyethylene glycol, and the artificial peptide comprises fluorescein, an LTVSPWY-PLGVR peptide fragment, 4-(Fmoc-aminomethyl) phenoxy butyric acid and a lauric acid alkyl chain which are connected in sequence. The preparation method comprises the following steps: preparing a film; preparing a core; the shell adsorbs the surface of the core and the membrane wraps the core-shell nanoparticles. The nano drug delivery system can realize quadruple synergy of chemotherapy, chemical kinetics, ferroptosis and immunogenicity death in the same particle, and breaks through the bottlenecks of high toxicity, fast drug resistance and weak immune activation of the traditional breast cancer therapy.

A phosphorous compound such as STMP is used as a cross-linking agent while making a starch nanoparticle in an emulsion process. Negative charge of the nanoparticle is reduced or reversed by adding cations and / or cationizing the starch optionally while forming the nanoparticles. Anionic active agents, such as fluoride or fluorescein, are optionally incorporated into the nanoparticle during the formation process. For example, a fluoride salt can also be used, which promotes the crosslinking reaction while also providing fluoride in the nanoparticle. The retention of both calcium and fluoride in the nanoparticle is improved when both salts are used. Alternatively, the nanoparticle may be used without added calcium and / or fluoride. The nanoparticles may be useful for tooth remineralization, the treatment of dentinal hypersensitivity, to treat caries, or as a diagnostic agent to locate carious lesions.

This invention relates to a chemiluminescent immunoassay reagent for tacrolimus, its preparation, and detection method. The reagent comprises a first reagent, a second reagent, and a magnetic separation reagent. The first reagent is a buffer solution containing a fluorescein-labeled anti-tacrolimus monoclonal antibody at a pH of 6.0–9.0, with the concentration of the fluorescein-labeled anti-tacrolimus monoclonal antibody being 1.0–5.0 μg / mL. The second reagent is a buffer solution containing a tacrolimus-alkaline phosphatase conjugate at a pH of 6.0–9.0, with the concentration of the tacrolimus-alkaline phosphatase conjugate being 0.05–0.20 μg / mL. The magnetic separation reagent is a suspension of magnetic microparticles coated with the anti-fluorescein monoclonal antibody. This invention enables the quantitative detection of tacrolimus with lower cost and higher accuracy and precision.

The present invention relates to the technical field of molecular imaging probes, and more specifically to a fluorescent probe targeting the estrogen receptor, a preparation method, and applications thereof. The fluorescent probe targeting the estrogen receptor is composed of a fluorescein ester, an estrogen receptor recognition group, raloxifene, and an alkane chain. The preparation method comprises the following steps: Step S1, hydrolyzing or hydrogenolyzing the carbamate of raw material I to synthesize an amino intermediate II; Step S2, condensing the amino intermediate II with FITC to obtain a fluorescent probe targeting the estrogen receptor III. The fluorescent probe targeting the estrogen receptor in the present invention can avoid the effects of short emission wavelength and low signal-to-noise ratio during probe application, and can be better used to promote the clinical application of the fluorescent probe targeting the estrogen receptor.

The application discloses a platelet antibody detection and cross matching blocking chromatography kit and a detection method thereof, wherein the kit comprises a U-shaped microwell plate, a chromatography medium for separating platelet immune complexes from non-cell particle components under the action of centrifugal force, freeze-dried platelets, a platelet diluent, a platelet fluorescent staining solution comprising fluoresceinized platelet antibodies specifically labeled for platelets, biotin-labeled anti-human globulin for specifically binding with antibodies bound to the surface of the platelets, avidinized fluorescein for coupling with the biotin-labeled anti-human globulin through a biotin-avidin specific binding system, negative control serum / plasma and positive control serum / plasma. The application combines the blocking chromatography technology with a high-sensitivity flow immunofluorescence method to construct an efficient and accurate platelet antibody detection and cross matching system, which is high in specificity and sensitivity, simple in operation and stable in detection results.

The present invention belongs to the technical field of magnetic thermal therapy, and particularly relates to a PD-L1@Fe3O4@LPS tumor magnetic thermal therapy probe and its preparation method and application. The PD-L1@Fe3O4@LPS tumor magnetic thermal therapy probe comprises a nanocarrier, a PD-L1 polypeptide fluorescein conjugate and an RS09 polypeptide; the PD-L1 polypeptide fluorescein conjugate and the RS09 polypeptide are conjugated to the surface of the nanocarrier, and the nanocarrier is a carboxylated PEG-modified magnetic iron oxide nanoparticles PEG-COOH-Fe3O4. This PD-L1@Fe3O4@LPS tumor magnetic thermal therapy probe simultaneously has the functions of inhibiting the immune escape of tumor cells and enhancing the release of tumor-killing cytokines by immune cells, and significantly enhances the immune activation performance of SPIO on tumors in MHT.

The invention discloses a visual paper-based cancer miRNA marker detection kit, the kit comprises an integrally formed foldable T-shaped filter paper, and a DNA probe TEprobe, urease and a color developing agent dropwise added on the filter paper, and the principle of the kit is as follows: the TEprobe is combined with a cancer nucleic acid marker (miRNA) to release Ag < + > to inhibit the activity of the urease; the composite signal response of the pH response probe phenol red and fluorescein is triggered, and the detection sensitivity is as low as 10-12 M. The kit provided by the invention can significantly shorten the detection time and reduce the sample demand, and realizes zero false positive rate and 97.10% specificity by combining with machine learning algorithm classification during result analysis. The performance is successfully verified in real samples of prostatic cancer, liver cancer and colorectal cancer, and a cheap and convenient means is expected to be provided for family self-inspection and cancer screening and diagnosis in resource-deficient areas.

A colorectal cancer related serum metabolite joint detection kit comprises a rectangular flat box body and four pieces of chromatography test paper, the test paper is arranged at intervals in the left-right direction, and each piece of test paper is formed by sequentially connecting a sample pad, a combination pad, a nitrocellulose membrane detection pad and an absorption pad. A detection line and a quality control line are arranged on the detection pad, the detection line is respectively coated with fluorescein-coated antibodies of lactic acid, 5 (S)-hydroxyeicosatetraenoic acid, lysophosphatidylcholine and diacylglycerol, and the quality control line is coated with streptomycete antibiotic protein-HRP. The kit body is provided with a serum sample adding hole and an observation window, so that sample loading and result reading are facilitated. By optimizing the antibody coating amount (1.7-2.3 [mu] g / cm < 2 >), the detection sensitivity and specificity are ensured, the color is developed only when the concentration of the target metabolite exceeds a threshold value, and the method has the characteristics of high accuracy and rapid detection.

The invention discloses a method for detecting peripheral blood immune cells through multi-parameter flow cytometry. The core of the method is that an optimized eight-color antibody combination is adopted and comprises CD45, CD3, CD19, CD16, CD56, CD4, CD8 and CD14, and a specific fluorescein pairing scheme is specified. According to the method, the detection stability is ensured through standardized sample dyeing and cracking washing processes in combination with precision calibration of a flow cytometer. In the data analysis stage, a set of serialization gate setting strategy is innovatively adopted, and fragments, adhesions and monocytes are sequentially eliminated, so that T cells (CD4 + and CD8 +) and B cells are accurately distinguished in a purified lymphocyte population, and NK cells are further subdivided into three functional subgroups, namely CD16 + CD56-, CD16 + CD56dim and CD16-CD56light. The method solves the problems of rough typing, poor accuracy and low repeatability in the prior art, and realizes efficient, accurate and repeatable detection of peripheral blood immune cells.

The invention belongs to the technical field of genetic breeding, and particularly relates to application of a DNA (deoxyribonucleic acid) fragment to regulation and control of GHR (growth hormone receptor) gene expression quantity and bovine body size, and a base sequence of the DNA fragment is shown as SEQ ID NO.12. According to the invention, SV detection and typing are carried out on individual genome sequencing data from different cattle varieties in the world, SV loci having significant selection in two differential populations are screened by a comparative genomics method, and SV variation located in a first intron region of a growth hormone receptor (GHR) is screened by combining gene functions annotated by SV, so that the SV variation is obtained. The influence of the SV on GHR expression is proved through analysis experiments such as a dual-fluorescein report system, it is proved that the SV can serve as a candidate molecular marker influencing the body type size, and an important target is provided for molecular breeding of beef cattle.

The present invention relates to methods of using CAR T cells. The present disclosure relates to methods of treating a patient having cancer by administering to the patient a composition comprising CAR T cells, and administering to the patient a small molecule linked to a targeting moiety by a linker, where the CAR T cells comprise a CAR and the CAR comprises an E2 anti-fluorescein antibody fragment. The present disclosure also relates to compositions for use in such methods.

Neuroendocrine tumors (NETs) are a class of highly heterogeneous tumors derived from neuroendocrine cells. Synaptic vesicle 2 (SV2) protein is a novel marker of NETs, and there are three subtypes of SV2A, SV2B and SV2C, wherein the structure of SV2C is more stable than the other two, and the largest cavity ring L4 thereof is the most important binding area with toxins. The present application obtains a nanobody of the extracellular soluble part L4 of SV2 by screening through phage display technology, and the nanobody labeled with a fluorescein, a small molecule of a chemical toxin or a nuclide is used to prepare a probe molecule, and the result can be specifically combined with neuroendocrine tumor cells. The present application provides a nanobody of the extracellular soluble part L4 of SV2, and the nanobody can be applied in the preparation of diagnostic and therapeutic preparations for neuroendocrine tumors.

The application provides an antibody composition for detecting acute T lymphoblastic leukemia by full-spectrum flow cytometry, a detection system and application thereof, and relates to the technical field of blood disease detection. The antibody composition is two groups of antibodies. The first group is 14 kinds of fluorescein-labeled cell membrane monoclonal antibodies, including anti-CD45, anti-CD99, anti-CD34, anti-CD48, anti-CD2, anti-CD3, anti-CD4, anti-CD5, anti-CD7, anti-CD8a, anti-CD16, anti-CD56, anti-CD94 and anti-TCR gamma delta. The second group is two kinds of intracellular monoclonal antibodies, including anti-nucleus TdT and anti-cytoplasm CD3. The two groups of antibodies are added into the same tube in sequence. The application also provides a flow cytometry sample for detecting acute T lymphoblastic leukemia based on full-spectrum flow cytometry, and a detection analysis method and system, so that the missed diagnosis rate and misdiagnosis rate of T-ALL detection are reduced.

Lipid-fluorescein conjugates, the compositions comprising same, and the methods of synthesis and use, such as in the treatment of cancer, are disclosed.

A phosphorous compound such as STMP is used as a cross-linking agent while making a starch nanoparticle in an emulsion process. Negative charge of the nanoparticle is reduced or reversed by adding cations and / or cationizing the starch optionally while forming the nanoparticles. Anionic active agents, such as fluoride or fluorescein, are optionally incorporated into the nanoparticle during the formation process. For example, a fluoride salt can also be used, which promotes the crosslinking reaction while also providing fluoride in the nanoparticle. The retention of both calcium and fluoride in the nanoparticle is improved when both salts are used. Alternatively, the nanoparticle may be used without added calcium and / or fluoride. The nanoparticles may be useful for tooth remineralization, the treatment of dentinal hypersensitivity, to treat caries, or as a diagnostic agent to locate carious lesions.

The present invention contemplates a method of treating a mammalian skin wound that extends at least into the epidermal layer of the skin of a subject mammal that comprises treating the wound in the substantial absence of actinic light by topical application of an aqueous pharmaceutical composition containing a wound closure-assisting amount of rose bengal, a lactone, salt, ester or amide hereof dissolved or dispersed therein as well as gel-inducing amount of a thickening agent (gellant) that causes the aqueous pharmaceutical composition to gel at a temperature of about 33° to about 40° C. The treated wound is thereafter covered with a dressing that is opaque to actinic light at least in the area over the wound. This treatment method is repeated a plurality of times over the following up to about fifteen days or until the wound is closed, whichever time of time is shorter.

The invention relates to the technical field of medical detection, in particular to an antibody composition and application. The antibody composition is composed of a CD33 antibody, an HLA-DR antibody, a CD14 antibody, a CD16 antibody and a CD45 antibody. The invention also provides an application of the antibody composition in detection of a peripheral blood mononuclear cell subset. The invention also provides a peripheral blood mononuclear cell subset determination method, which comprises: adding the fluorescein-labeled antibody composition and PBS to a blood sample to be detected, incubating in a dark place, adding the PBS, washing, centrifuging, discarding the supernatant, adding hemolysin, standing in a dark place, adding the PBS, washing, centrifuging, discarding the supernatant, adding the PBS, and detecting the peripheral blood mononuclear cell subset. And performing flow cytometry on a machine for detection. The invention solves the problem that the false positive result is easily caused when the existing antibody combination is used for detecting the mononuclear cell subpopulation.

Some embodiments of the methods and compositions provided herein include therapies for treating, ameliorating, or inhibiting cancer (e.g., osteosarcoma) in a human subject. Some embodiments include administering to a subject a fluorescein isothiocyanate (FITC)-folate conjugate, or a pharmaceutically acceptable salt thereof, in an incremental dose regimen following or concurrently with administering to the subject a chimeric antigen receptor (CAR) T cell therapy (e.g., T cells comprising anti-fluorescein CAR). In some such embodiments, the dose increment regimen includes a dose increment period to confirm a maximum tolerable dose (MTD) of the FITC-folate conjugate, or a pharmaceutically acceptable salt thereof.

The invention belongs to the technical field of biological medicine, and discloses application of a chick embryo allantoic membrane model in lung adenocarcinoma drug resistance mechanism research. The chick embryo allantoic membrane model is obtained by hatching fertilized white-shell eggs in a bird incubator at 37.5 DEG C and 65% humidity. After cancer cells are transplanted to Y-shaped blood vessels of the chorioallantoic membrane, H2228 cells continuously expressing Venus fluorescent protein are transplanted to the chorioallantoic membrane so as to confirm the implantation condition. Before and after medicine injection, all organs of the cut tumor and embryo are shot under fluorescence, and the fluorescence intensity of all the organs is quantitatively analyzed through a fluorescein filter. According to the invention, a chick embryo allantoic membrane model is used for verifying the cancer inhibition effect of Chb-M'in ALK mutant lung adenocarcinoma cells for the first time, the experiment cost is relatively low, and the experiment period is relatively short.

The present invention contemplates a method of treating a mammalian skin wound that extends at least into the epidermal layer of the skin of a subj ect mammal that comprises treating the wound in the substantial absence of actinic light by topical application of an aqueous pharmaceutical composition containing a wound closure-assisting amount of rose bengal, a lactone, salt, ester or amide hereof dissolved or dispersed therein as well as gel-inducing amount of a thickening agent (gellant ) that causes the aqueous pharmaceutical composition to gel at a temperature of about 33° to about 40° C. The treated wound is thereafter covered with a dressing that is opaque to actinic light at least in the area over the wound. This treatment method is repeated a plurality of times over the following up to about fifteen days or until the wound is closed, whichever time of time is shorter.

This invention relates to the field of nitric oxide donor technology, and particularly to a fluorescein-fused nitric oxide donor, its preparation method, and its application. The method includes the following steps: adding N,N-dimethylformamide to a mixture of an aromatic compound containing a para-active halogenated side chain and potassium carbonate; stirring; adding 3-trifluoromethyl-4-nitroaniline; quenching the reaction with saturated brine; drying and vacuum concentration; and purifying. Then, mixing fluorescein, potassium carbonate, and dimethyl sulfoxide and stirring; placing the first mixture in an ice-water bath; adding a second mixture; quenching the reaction with saturated brine; drying and vacuum concentration; and purifying to obtain the fluorescein-fused nitric oxide donor. This constructs a functional molecule that combines NO release function and fluorescence response characteristics, achieving an integrated release-response-imaging function. This solves the problem that existing NO donors are difficult to monitor in real time, making it impossible to intuitively and in real-time trace NO release behavior, thus limiting the practical application of nitric oxide donors.

The invention relates to the technical field of medical image processing and disease auxiliary diagnosis, in particular to a diabetic neuropathy auxiliary detection system based on ophthalmologic images. According to the system, the fundus image acquisition module, the hysteresis analysis module, the region fusion module, the microhemangioma influence degree acquisition module and the diabetic neuropathy detection module are arranged to analyze gray change characteristics of pixel points in an FFA image, the hysteresis degree of the pixel points filled with fluorescein is acquired, region fusion is carried out according to the hysteresis degree, and the detection accuracy of the diabetic neuropathy is improved. And quantifying the influence degree of the microhemangioma according to the average lag degree of the fusion region and the region area to realize the judgment of the diabetic neuropathy. The system obtains the influence degree of the microhemangioma by quantitatively analyzing the filling and diffusion rules of fluorescein in the microhemangioma and the surrounding area of the microhemangioma, and achieves the auxiliary detection of the diabetic neuropathy, thereby assisting doctors to efficiently and accurately detect the diabetic neuropathy, and effectively improving the diagnosis efficiency and accuracy.

Disclosed herein are pharmaceutical compositions comprising (a) a fluorescein labeled nerve targeting peptide of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate or isotope thereof: (I) (b) an additive comprising a pH regulator, a buffer, a filler, a tonicity regulator, a preservative, or any combination thereof; and (c) an aqueous vehicle wherein the pH of the composition is from about 7 to about 9 and the concentration of the peptide in the aqueous composition is from about 25 mg / mL to about 200 mg / mL based on the total volume of the aqueous composition. Also disclosed are methods for making and using the pharmaceutical compositions described herein.