113 results about "Affinity Reagent" patented technology

A method for analysing biological sample is provided, comprising the following steps: a) providing a biological sample with a plurality of target analytes (106, 306); b) introducing at least a first marker (108, 124, 202, 308, 400, 500, 502, 504) into the biological sample, the first marker comprising a first affinity reagent (110, 204, 310, 506) and a first optically detectable label (112, 126) bound to the first affinity reagent (110, 204, 310, 506), wherein the first affinity reagent (110, 204, 310, 506) is configured to specifically bind to one of the target analytes (106, 306) and configured to be degraded by a degradation agent (120, 200); c) generating a first optical readout of the biological sample with the first marker; d) applying the degradation agent to degrade at least the first affinity reagent (110, 204, 310, 506); e) introducing at least a second marker (108, 124, 202, 308, 400, 500, 502, 504) into the biological sample, the second marker comprising a second affinity reagent (110, 204, 310, 506) and a second optically detectable label (112, 126) bound to the second affinity reagent (110, 204, 310, 506), wherein the second affinity reagent (110, 204, 310, 506) is configured to specifically bind to one of the target analytes (106,112) and configured to be degraded by the degradation agent (120, 200); f) generating a second optical readout of the biological sample with the second marker.

The present disclosure provides a method of processing an analyte, including steps of (a) providing an analyte including an epitope and a docker; (b) providing an affinity reagent, wherein the affinity reagent includes a paratope that recognizes the epitope and a tether that recognizes the docker; and (c) contacting the analyte with the affinity reagent, whereby the affinity reagent associates with the analyte via binding of the paratope to the epitope and via binding of the tether to the docker. Optionally, the method further includes the step of (d) detecting association of the affinity reagent with the analyte, thereby identifying the analyte.

The present disclosure provides methods and systems for performing single-molecule detection using fabricated integrated on-chip devices. Provided herein is a composition, comprising: (a) a substrate; (b) a structured nucleic acid particle coupled to the substrate, wherein a biological entity is coupled to the structured nucleic acid particle; and (c) a fluorescent detection agent, wherein the fluorescent detection agent comprises an affinity reagent attached to another structured nucleic acid particle, wherein the affinity reagent is bound to the biological entity.

Aptamers as nucleic acid affinity reagents are gaining traction in analytical applications owing to their high stability, compact size, and amenability to chemical modifications. It is highly desired to generate aptamers with different binding affinities to meet the requirements of various applications. However, systematic evolution of ligands by exponential enrichment (SELEX), the gold standard for aptamer generation, is unable to quantitatively produce aptamers with desired binding affinities and requires multiple rounds of selection to eliminate false positive hits. Here we introduce Pro-SELEX, a new approach for rapid discovery of aptamers with precisely defined binding affinities that combines efficient particle display, high-performance microfluidic sorting and high-content bioinformatics. Using the Pro-SELEX workflow, we were able to investigate the binding performance of individual aptamer candidates under different selective pressure in a single round of selection. Using human myeloperoxidase (MPO) as a target, we demonstrated that aptamers with dissociation constants spanning a twenty-fold range of affinities can be identified within one round of Pro-SELEX. Our selection strategy will have a significant impact on the fields of biosensing and bioseparation by generating aptamers with tailored binding affinities.

A method for analysing a biological sample includes providing a biological sample with a plurality of target analytes, and introducing a first marker into the biological sample. The first marker includes a first affinity reagent and a first optically detectable label bound to the first affinity reagent. The first affinity reagent is configured to specifically bind to one of the target analytes. The method further includes generating a first optical readout of the biological sample with the first marker, applying the degradation agent to degrade the first affinity reagent, and introducing a second marker into the biological sample. The second marker includes a second affinity reagent and a second optically detectable label bound to the second affinity reagent. The second affinity reagent is configured to specifically bind to one of the target analytes. The method further includes generating a second optical readout of the biological sample with the second marker.

Systems and methods for obtaining qualitative or quantitative measurements of proteoforms of polypeptides are described. The described methods include measurements of affinity reagent binding on single-molecule polypeptide arrays to distinguish between polypeptide isoforms. The described methods may provide high resolution quantitative comparisons of proteoforms with very low copy numbers.

An affinity reagent for analysing a biological sample includes a nucleic acid backbone, and a barcode oligonucleotide attached to the nucleic acid backbone. The nucleic acid backbone is configured to specifically bind to a target analyte by a complex structure thereof. The nucleic acid backbone is configured to maintain the complex structure in presence of the barcode oligonucleotide.

An in vitro method for determining basophil activation in a sample is provided. Other aspects relate to determining an efficacy of an allergen-specific desensitization treatment of a subject. The method includes providing a sample from the subject, treating the sample with an allergen and determining a level of at least one basophil inhibitory marker and at least one basophil activation marker in the sample, the basophil inhibitory marker includes CD32. Corresponding systems and machine learning methods are also described. The use of an affinity reagent that specifically binds CD32 in an in vitro method for determining basophil activation is also provided, for example in the context of determining an efficacy of an allergen-specific desensitization treatment of a subject. A kit and corresponding uses of affinity reagents is also detailed.

Disclosed herein are chromatographic materials comprising crosslinked streptavidin molecules. The crosslinked streptavidin chromatographic materials may be used to prepare affinity chromatographic materials using biotinylated affinity agents, such as a biotinylated antibody, biotinylated antigen-binding fragment, or biotinylated oligonucleotide. Due to the crosslinked nature of the streptavidin molecules, the materials provided herein have reduced to no streptavidin leachate, resulting in improved performance.

Single-cell multi-omics by co-encapsulating a single cell with two beads, the first an RNA barcoding bead having barcoded mRNA capture primer oligonucleotides attached on the bead surface; and the second a DNA barcoding bead having two types of oligonucleotides releasably attached to the surface: (1) barcoded adapter oligonucleotides that are complementary to oligonucleotides bound to the transposase that are eventually incorporated into gDNA fragments and (2) polyadenylated barcoded oligonucleotides containing the same barcode sequence as the adapters. In addition, integrated analysis of RNA and protein, including intracellular protein, from individual cells using similar co-encapsulation of a single cell, an RNA barcoding bead, and with / without a specific or non-specific protein binding bead in a microwell, to avoid protein fixation by first lysing the cell to liberate intracellular contents, and then capturing protein either on a solid surface or in solution with barcoded affinity reagents.

Disclosed is a method for detecting analytes in a biological sample, comprising following steps: a. contacting the biological sample with at least a first marker (100a) and at least a second marker (100b), wherein the at least first marker comprises a first affinity reagent (108a) configured to bind to a first analyte (102a) and a first nucleic acid label (104a) and wherein the at least second marker comprises a second affinity reagent (108b) configured to bind to a second analyte (102b) and a second nucleic acid label (104b), b. incubating the biological sample with said first marker and said second marker, in particular for a sufficient amount of time, to allow the at least first affinity reagent (108a) to bind to the first analyte (102a) and to allow the at least second affinity reagent (108b) to bind to the second analyte (102b), and c. adding at least one bridging strand (200) configured to bind to the first (104a) and second nucleic acid label (104b).

Disclosed herein are improved in situ methods for mapping the location of a protein on chromatin and DNA-based in situ methods for measuring transcription in a cell from a formalin-fixed paraffin-embedded (FFPE) sample. The methods can include steps of treating the FFPE sample to remove paraffin; permeabilizing the sample; contacting the sample with a first affinity reagent that specifically binds to a targeted chromatin protein or a protein involved in transcription regulation, wherein the first affinity reagent is coupled to a transposome comprising at least one transposase and a transposon; activating the at least one transposase under low ionic conditions, thereby cleaving and tagging chromatin DNA; excising the tagged DNA segment associated with the targeted chromatin protein or protein involved in transcription regulation; and determining the nucleotide sequence of the excised tagged DNA segment, thereby mapping the genomic location of the targeted protein on chromatin or transcriptional activity on chromatin.

The present disclosure provides a method of processing an analyte, including steps of (a) providing an analyte including an epitope and a docker; (b) providing an affinity reagent, wherein the affinity reagent includes a paratope that recognizes the epitope and a tether that recognizes the docker; and (c) contacting the analyte with the affinity reagent, whereby the affinity reagent associates with the analyte via binding of the paratope to the epitope and via binding of the tether to the docker. Optionally, the method further includes the step of (d) detecting association of the affinity reagent with the analyte, thereby identifying the analyte.

The invention discloses a novel high-sensitivity and high-flux carbohydrate compound recognition method based on chlorine enhanced ionization. The method comprises the following steps: extracting carbohydrates in a to-be-detected sample to obtain a carbohydrate extracting solution; chloromethyl triphenylphosphonium chloride is optimally selected from 23 quaternary phosphonium salts, quaternary ammonium salts and inorganic ammonium salts as a chlorine affinity reagent added to a mobile phase, and a carbohydrate extracting solution is subjected to targeted and non-targeted scanning analysis by liquid chromatography-tandem high-resolution mass spectrometry (UPLC-ESI-HRMS); and identifying unknown carbohydrate compounds and analyzing the content of known carbohydrates by utilizing a high-resolution mass spectrum scanning result. According to the method, the chloromethyl triphenylphosphonium chloride chloroaffinity reagent is added into the mobile phase, so that the detection sensitivity and the separation effect of the carbohydrates are remarkably improved, and the carbohydrates are analyzed with small sample amount, low interference and high sensitivity; meanwhile, the chlorine isotope labeled carbohydrates have the isotope distribution characteristic, and polysaccharide can generate double-charge molecular ion peaks, so that the mass spectrum detection mass range is remarkably increased, a new technology is provided for screening the high-quality coverage range of the carbohydrates in food, biological and environmental samples, and the method has originality.

A marker for analysing a biological sample includes an affinity reagent, at least one labelling moiety, and at least one linker oligonucleotide. The at least one linker oligonucleotide is attached to the affinity reagent, and the at least one labelling moiety is attached to the at least one linker oligonucleotide.

Methods and systems are provided for accurate and efficient identification and quantification of proteins. In an aspect, disclosed herein is a method for iteratively identifying candidate proteins within a sample of unknown proteins, the method comprising receiving information of binding measurements of each of a plurality of affinity reagent probes to the unknown proteins, each affinity reagent probe configured to selectively bind to one or more candidate proteins; comparing at least a portion of the information of binding measurements against a database comprising a plurality of protein sequences, each protein sequence corresponding to a candidate protein; and iteratively generating a probability that each of one or more candidate proteins is present in the sample based on the comparison of the information of binding measurements of the candidate proteins against the database comprising the plurality of protein sequences.

The invention discloses a method for synthesizing chiral morpholine from propargyl alcohol ester. Chiral morpholine is an important drug molecular skeleton and is widely applied to the aspects of analgesia, anesthesia, tumor resistance, heart treatment and the like as a drug active ingredient, but an efficient synthesis method is lacked. According to the method disclosed by the invention, under the regulation and control of arylboronic acid and a specific chiral ligand, propargyl alcohol ester and an affinity reagent react to obtain optically pure (87-97% ee) morpholine. The method has the characteristics that the catalyst and the ligand are wide in source, and the substrate is cheap and easy to obtain; the chiral morpholine has excellent enantioselectivity and good functional group compatibility, can be suitable for structural modification of bioactive molecules and intermediates, and is a simple and efficient asymmetric catalysis strategy for synthesizing chiral morpholine.

Provided herein are techniques relating to the identification of binding sites of DNA binding proteins, and in particular, but not exclusively, to methods, systems and kits for simultaneous mapping of binding sites of multiple proteins in the same cell using affinity reagent specific barcodes.

Systems and methods for obtaining qualitative or quantitative measurements of proteoforms of polypeptides are described. The described methods include measurements of affinity reagent binding on single-molecule polypeptide arrays to distinguish between polypeptide isoforms. The described methods may provide high resolution quantitative comparisons of proteoforms with very low copy numbers.

In various aspects, the present disclosure relates to an adsorbent for separating at least one target protein from a liquid sample, the adsorbent comprising a solid support comprising at least one attached high affinity reagent having affinity for the at least one target protein. Other aspects of the present disclosure include kits containing such adsorbents and methods of processing samples using the kits.

The present invention relates to affinity reagents that are capable of specifically binding to a surface antigen of a virus belonging to the family of human endogenous retroviruses (HERV), in particular to a HERV envelope protein, for use in the prophylactic and / or therapeutic treatment of a cancer in a subject in need thereof. The invention relates also to the use of such affinity reagents in the diagnosis or prognosis of cancer, in particular lymphoma.

Provided herein are nanobodies, polypeptides comprising the same, and uses thereof. The nanobody's variable region includes 3 complementarity determining regions (CDRs) and framework regions (FRs), wherein the CDRs are as follows: CDR1: Ser-Gly-Xaa11-Xaa12-Phe-Xaa13-Xaa14-Asn-Xaa15 (Formula I); CDR2: Xaa21-Thr-Xaa22-Xaa23-Gly-Xaa24-Thr (Formula II); CDR3: His-Xaa31-Asp-Glu-Xaa32-Arg-Xaa33-Ser-Xaa34-Trp-Thr-Thr-Ser-Asn-Xaa35 (Formula III). The nanobodies and their polypeptides exhibit high affinity and activity, specifically recognizing and binding AAV. Affinity agents prepared therefrom have strong AAV adsorption capacity, suitable for AAV affinity chromatography to facilitate its industrial application. They are also applicable to AAV detection, enabling simultaneous detection of empty capsids and viral particles.

The present disclosure relates to the fields of life science and medicine. Specifically, the present disclosure relates to a method for culturing, detecting and screening a cell. A method for capturing a cell secretion comprises: performing culture treatment on a single cell under a suitable condition; and performing controllable rotary flotation treatment on a first affinity reagent in a single cell culture treatment system, wherein the affinity reagent can specifically bind to the secretion of the single cell.

This invention belongs to the field of rapid detection technology, specifically relating to a rapid, real-time detection method for influenza A based on electrochemical biosensors. This invention provides an aptamer that specifically recognizes the influenza A virus, and uses a pairing affinity reagent to enrich the aptamer onto magnetic beads to prepare a probe that specifically recognizes the influenza A virus. Then, using this probe, MB solution, and screen-printed electrodes, an electrochemical biosensor that specifically recognizes the influenza A virus is fabricated. Qualitative or quantitative detection of influenza A virus using the electrochemical biosensor of this invention eliminates the need for complex pretreatment and viral nucleic acid amplification, greatly simplifying the operation. The results are stable and reliable, making it suitable for real-time detection applications.

Compositions and methods for isolating and detecting nucleic acid in a biological sample are provided. The compositions and methods utilize a solid support and an affinity reagent for isolating and detecting the nucleic acid.

A method of identifying extant proteins, including (a) inputting to a computer processor: (i) a plurality of empirical binding profiles, individual empirical binding profiles including empirical binding outcomes for binding of an extant protein to a plurality of different affinity reagents, (ii) a plurality of candidate outcome profiles, individual candidate outcome profiles including binding outcomes for binding of a candidate protein to the plurality of different affinity reagents, and (iii) a plurality of pseudo outcome profiles, individual pseudo outcome profiles including a rearrangement of a candidate outcome profile; (b) performing a process in the computer processor to identify extant proteins based on the empirical binding profiles of the extant proteins and the plurality of candidate outcome profiles; and (c) performing a process in the computer processor to determine a false discovery statistic for the extant proteins based on the plurality of pseudo outcome profiles.

The present disclosure provides a method of identifying a polypeptide. The method can include steps of (a) attaching a polypeptide to a particle or solid support, thereby producing an immobilized polypeptide having a plurality of amino acids linked to the particle or solid support; (b) fragmenting the immobilized polypeptide, whereby the particle is attached to a set of fragments of the polypeptide; (c) performing a binding assay including contacting the set of fragments with a plurality of affinity reagents and detecting binding of affinity reagents of the plurality of affinity reagents to the set of fragments; and (d) identifying the polypeptide from results of the binding assay

A method for producing a purified polyclonal anti-HCP antibody preparation is disclosed. The method can comprise acquiring a sample comprising antibodies from an animal immunized with HCP of a host cell, separating antibodies from the sample by contacting the sample with a protein A affinity reagent to provide an antibody preparation, separating anti-HCP antibody from the antibody preparation by contacting the antibody preparation with an HCP-affinity reagent comprising HCP coupled to a substrate, and isolating the separated polyclonal anti-HCP antibody preparation.

In various aspects, the present disclosure relates to an adsorbent for separating at least one target protein from a liquid sample, the adsorbent comprising a solid support comprising at least one attached high affinity reagent having an attachment with affinity to the at least one target protein. Other aspects of the present disclosure include kits containing such sorbents and methods of processing samples using the kits.