33 results about "Affinity labeling" patented technology

Method of enriching specific cells from cellular samples are disclosed, comprising contacting in solution a cellular sample with affinity-tagged ligands (ATLs) each comprising a first ligand linked to an affinity tag, wherein the ligand selectively binds a cellular marker of the rare cells and the affinity tag can be selectively captured by a capture moiety, wherein the affinity tags do not comprise a magnetic particle; and flowing the sample through a microfluidic device comprising the capture moiety to selectively retain ATL-bound cells. Methods for enriching circulating tumor cells, and devices for enriching specific cells from cellular samples are also disclosed.

Novel methods and compositions are provided for preferentially bonding an oligomeric molecule to a macromolecular target where the target is a member of a complex mixture and/or there is preferential bonding at one of a plurality of available bonding sites of the macromolecular target. The methods represent a complete system for both producing and identifying affinity label molecules from a combinatorial library which preferentially bind to a marcomolecular target or target site and preferentially binding those affinity labels to a macromolecule of interest either ex vivo or in vivo. Macromolecular targets include a variety of cellular- and non-cellular-associated proteins.

Methods and compositions are provided for identifying compounds having affinity or complementarity to a target molecule. Compounds according to the invention may be described by the formula E-C_a—R—C_b-A, wherein E is a therapeutic or diagnostic agent, R is a reactive group, C_a and C_b are connector groups between E and R and between R and A, respectively, and A is an affinity group comprising the sequence F-1-Y-E-E. Compounds according to the invention may be used for labeling the target molecule, particularly where the target molecule is naturally found in a complex mixture, such as a physiological fluid, like blood. By affinity labeling in vivo, the lifetime of physiologically active entities can be greatly enhanced by becoming bound to long-lived blood components. The covalently bound entity may also serve as an antagonist or agonist of a particular binding protein or as an enzyme inhibitor.

The present invention provides methods, kits, and compositions for purifying HDL molecules from a sample (e.g., blood sample) using HDL tagging molecules comprising an HDL lipophilic core binding peptide (e.g., portion of ApoA1) and an affinity tag. The present invention also provides methods, kits, and compositions for detecting non-fragmented ApoA1 with mass spectrometry. The present invention further provides methods, kits, and compositions for tagging HDL molecules in a sample with detectably labeled ApoA1 molecules such that the ratio of detectably labeled ApoA1 molecules to native ApoA1 proteins may be determined.

The invention is related to a method and test kits for quantitative determination of polynucleotide amounts present in a sample. The test kit comprises organized pools with polynucleotide probes having distinct sizes and optionally provided with tracer tags or primer tags. The probes are allowed to hybridize with affinity tagged analyte polynucleotides from the sample. The result is hybrids, which can be recovered on a separation aiding tool provided with the pair of the affinity tag. After the quantitative release of the probes, the probes are either directly recorded, or if primer tagged, they are amplified and optionally provided with a tracer tag before recording. The invention provides a sensitive and quantitative determination of the amount polynucleotides present in a cell or tissue sample and allows a quantitative assessment of variations in the amounts of polynucleotides as a response to inherent changes or due to external stimuli.

The invention relates to a method and test kit for quantitative determination of the amounts or relative proportions of polynucleotides in a mixture. The invention enables assessment of dynamic variations in a mixed population of organisms using affinity aided solution hybridization. The test kit comprises organized pools of polynucleotide probes having approximately the same number of nucleotides, which are distinguishable using resolution enabling tags providing the probes with different sizes. The resolution enabling tags may simultaneously act as tracer, affinity or primer tags. The probes are allowed to hybridize with affinity tagged analyte polynucleotides. The result is hybrids, recoverable on separation aiding tools provided with counterparts of the affinity tag. After the quantitative release of the probes, the individual probes can be amplified and recorded. The method and test kit are useful for determining hygienic and epidemiologic situations and evaluating the effect of antibiotic treatment and sanitary measures.

The invention discloses an isoandrographolide photoaffinity labeling molecular probe. The structure of the molecular probe is represented in a formula (I), wherein n1 ranges from 1 to 5 and n2 ranges from 0 to 2. Furthermore, the invention discloses a preparation and a pharmaceutical composition of the isoandrographolide photoaffinity labeling molecular probe. The compound, which is excellent in TNF-alpha inhibitory activity, can be used for researching and discovering target protein of isoandrographolide having an action of regulating macrophage function, and the compound, as a tool medicine, can be used for medicine development of a diagnostic reagent of the target protein as well as a regulator thereof.

The invention discloses a method for purifying a nano-antibody drug by employing Protein A affinity chromatography. The method comprises the steps of: (1) adjusting the pH of a nano-antibody sample; (2) equilibrating the chromatography column with an equilibration buffer solution; (3) loading a sample adjusted by the step (1) to the chromatography column; (4) washing the product with a washing buffer solution; and (5) eluting the material with an elution buffer solution. The method provided by the invention has the advantages of simple operation, affinity purification without additional affinity labeling, higher purity and easy process amplification.

The invention relates to a method for analyzing a genomic region of an organism, comprising four major parts. The first part involves the isolation of mRNA from a selected organism that is used for the preparation of small single stranded DNA fragments with one adaptor containing an affinity label. These DNA fragments are used in part three. In the second part, genomic DNA from the same or a related organism is isolated. This genomic DNA is fragmented and ligated to adaptor molecules. In the third part, these genomic fragments are hybridized with single stranded DNA fragments from part one, and the hybrids formed in this process are used for synthesis of DNA fragments. These fragments will be used in part four which involves sequencing of these fragments using one of the available high throughput sequencing methods.

The invention provides a sequencing method based on a gene capture technology. The sequencing method has the principle that a recombinase compound is used for assisting a single-stranded DNA probe with a closed 3' end to efficiently and specifically hybridize into a double-stranded target DNA molecule under the condition of 37 DEG C, and then an affinity label on the single-stranded DNA probe is used for carrying out specific enrichment on the target DNA. The method not only has the advantages of relatively strong long probe pairing, low probe design requirement and the like of a probe hybridization capture technology, but also has the advantages of simplicity in operation, short time consumption, no need of thermal denaturation of DNA, high capture efficiency, low cost and the like of a CRISPR/dCas9 system, and the probe does not participate in subsequent amplification, so that the identification accuracy is improved. Therefore, compared with other capture technologies, the RATE-seq has the obvious advantages of simplicity in operation, high capture efficiency, extremely short consumed time (30 minutes), good stability, low cost and the like, and is very suitable for gene capture and automatic application. In addition, the RATE-seq can effectively separate and remove human host DNA in a pathological sample, and the detection rate of pathogenic microorganisms is increased.

An irreversible affinity labeling agent for labeling glutathione-S-transsulfurase sj(GST) comprises an N-(dimethylhydroquinone yloxyethyl)-(N-p-substituted amido benzoyl) glycine monovalent labeling agent, and a bisamide bivalence labeling agent which is derived by the N-(dimethylhydroquinone yloxyethyl)-(N-p-substituted amido benzoyl) glycine monovalent labeling agent and dialkyl triamine; the irreversible labeling agent includes four types of functional groups, namely a dimethylhydroquinone yloxyethyl hydrophobic orienting group for guiding the labeling agent to enter a hydrophobic site at a GST active center, a modifying functional base which is represented by alpha-halogenated acyl and reacts with sulfydryl/ amino close to the GST active center, an N-bis-substituted amino acid coupling functional base which is in amide coupling with micro-nano materials, and para aminobenzoic acid or glutamic acid or acylates of para aminobenzoic acid and glutamic acid, wherein the acylates of para aminobenzoic acid and glutamic acid can supply flexible arms required by the modifying functional group for a connecting skeleton. According to the affinity labeling agent, carboxyl/ amino of the coupling functional group is in amide coupling with the micro-nano material amino/ carboxyl, and then affinity labeling is performed on sjGST, thus achieving covalent linkage outside the active bit of fusion expression protein, and as a result, a novel protein functional micro-nano material can be prepared.

The present invention relates to kits and methods for the isotopic/isobaric labeling and the subsequent affinity selection and analysis of proteinaceous molecules. In particular, the invention relates to a kit-of-parts comprising a combinatorial plurality of labeling reagent for the labeling of proteinaceous molecules, each labeling reagent comprising an isobaric label component, an isotopic label component, and a reactive group capable of reacting with a proteinaceous molecule, wherein the isotopic label component concomitantly is an affinity tag. The invention is also directed to a corresponding method for the analysis of proteinaceous molecules, comprising labeling at least one subset of the proteinaceous molecules present by employing a kit-of-parts as defined herein and subsequently separating the labeled molecules by affinity purification via the affinity tag comprised in the label. Finally, the invention relates to the uses of such methods for protein expression profiling or proteomic analyses.

A modified Streptococcus aureus Cas9 (SaCas9) protein with a mutation at an N413 position, and optionally one or more of a nuclear localization sequence, a cell penetrating peptide sequence, an affinity tag and/or a fusion base editor protein, and a kit that comprises said modified protein. A method for altering the genome of a cell, the method including the step of using the modified protein of the invention.

The invention relates to a monomolecular mechanics method for measuring the accurate length of human telomere. The method comprises the following steps of: firstly, extracting genome DNA of a target cell; digesting the genome by using combined enzyme digestion, and reserving telomere DNA; carrying out affinity labeling on the two ends of telomere DNA by adopting digoxin and biotin respectively soas to obtain a telomere DNA structure capable of being used for Monomolecular mechanical measurement; secondly, fixing the telomere structure between streptavidin-coated magnetic beads and the glass surface paved with the anti-digoxin antibody, and carrying out a DNA mechanical tensile experiment on a monomolecular instrument; and finally, using a worm model to perform length conversion between nanometer and basic group pairs on the actually measured telomere length, so as to realize accurate measurement of the telomere length. The invention belongs to the field of precise instrument measurement and analysis and cell molecular biology, and develops a new method for directly measuring the accurate length of telomere for cell research, health screening and clinical drug therapeutic effect evaluation.

Synthetic macromolecular conjugate for selective interaction with proteins has a synthetic copolymer, and at least one binding group and at least one further group selected from an affinity tag and an imaging probe, and at least one binding group and at least one further group being bound via covalent bond to the synthetic copolymer. The macromolecular conjugate is suitable in particular for identification, visualization, quantification or isolation of proteins and/or cells.