Abstract: Provided herein are high-throughput, population-wide serotyping and antibody profiling assays. Disclosed variants of a Digital Serotyping assay employ next generation sequencing to measure the “serotyping profile” of barcoded subject serum antibodies tested against a range of DNA-tagged pathogen-derived antigens. The disclosed assay setup enables multiplexing in both the sample and antigen dimensions, generating a large multi-dimensional serotyping data set for more comprehensive serotyping profiling of large populations across a large number of antigens and possible pathogens. Moreover, the ability to easily scale and multiplex the number of peptide epitopes allows rapid updating of the assay content to monitor the ever-changing spectrum of pathogens. Additional applications of this technology include cancer immunology and autoimmune conditions (e.g., neoantigen or autoimmune profiling), screening for toxins, antibody therapeutics development, biosecurity, and veterinary medicine.

Abstract: Methods and compositions for quantitative immunoassays are provided, in which ligand-conjugated probes are used to label samples and ligand-surfaced microspheres are used as quantitative reference standards. Certain embodiments provide a method of quantitative flow cytometry where ligands are oligonucleotides, and a sample comprising one or more cells is contacted with a hybridized antibody::fluorophore labeled targeting construct to label the cells, and the labeled cells are analyzed. In some embodiments, a population of quantitative labeled oligospheres labeled with the same fluorescent label as the cells is analyzed using the flow cytometer and used to create a quantitative standard curve of cytometer intensity versus molecules fluorescent label per oligosphere event. A standard curve trendline is established and used to determine the molecules of fluorescent label per cellular event for the antigen-positive cell populations.

Abstract: Methods and compositions for quantitative immunoassays are provided, in which ligand-conjugated probes are used to label samples and ligand-surfaced microspheres are used as quantitative reference standards. Certain embodiments provide a method of quantitative flow cytometry where ligands are oligonucleotides, and a sample comprising one or more cells is contacted with a hybridized antibody::fluorophore labeled targeting construct to label the cells, and the labeled cells are analyzed. In some embodiments, a population of quantitative labeled oligospheres labeled with the same fluorescent label as the cells is analyzed using the flow cytometer and used to create a quantitative standard curve of cytometer intensity versus molecules fluorescent label per oligosphere event. A standard curve trendline is established and used to determine the molecules of fluorescent label per cellular event for the antigen-positive cell populations.

Abstract: A bead construct includes a bead, a fluorophore molecule, first and second linker molecules and a ligand molecule. The fluorophore molecule comprises at least two subunits connected to each other by at least one internal linker that provides conformational freedom to the at least two subunits. An optical signal emitted by the fluorophore molecule changes when reducing the conformational freedom of the at least two subunits to each other. The first linker molecule is connected to the bead and to one of the subunits of the flurophore molecule. The second linker molecule is connected to another one of the subunits of the fluorophore molecule. The ligand molecule is connected to the second linker molecule.

Abstract: The present invention relates to a method for verifying the integrity of biological source samples subjected to multistep bioassays that comprise massively parallel sequencing of the sample genomic nucleic acids. The integrity of the biological source samples is verified using unique marker nucleic acids that are combined with the biological source sample, and are sequenced concomitantly with the genomic nucleic acids of the biological source sample. The method provides verification of individual samples in single- and multiplex massively parallel sequencing assays.

Abstract: Provided herein is technology relating to next-generation sequencing and particularly, but not exclusively, to methods and compositions for preparing a next-generation sequencing library comprising short overlapping DNA fragments and using the library to sequence one or more target nucleic acids.

Abstract: Provided herein is technology relating to next-generation sequencing and particularly, but not exclusively, to methods and compositions for preparing a next-generation sequencing library comprising short overlapping DNA fragments and using the library to sequence one or more target nucleic acids.

Abstract: Methods and compositions for synthesizing nucleic acid sequences in an emulsion are provided.

Abstract: Provided is a set of mass labels, each mass label in the set comprising a mass marker moiety attached via a cleavable linker to a mass normalization moiety, each mass label in the set having a common mass; wherein the set comprises a plurality of groups of mass labels, the mass of the mass marker moiety being the same for mass labels within a group, the mass of the mass marker moiety being different between groups; the mass marker moiety is capable of fragmentation into two or three fragments; and the mass of at least one fragment of the mass marker moiety differs between mass labels within a group.

Abstract: The invention describes methods for labeling or detecting of immobilized poly(amino acids), including peptides, polypeptides and proteins, on membranes and other solid supports, using fluorescent dipyrrometheneboron difluoride dyes. Such immobilized poly(amino acids) are labeled or detected on blots or on arrays of poly(amino acids), or are attached to immobilized aptamers.

Abstract: The present invention relates to a method for absolute quantification of polypeptides.

Abstract: Provided is a method for characterizing a molecule by mass spectrometry, which molecule comprises one or more free amino groups, which method comprises: (a) reacting one or more free amino groups in the molecule with a mass tag reagent comprising a reactive functionality capable of reacting with an amino group, and a tertiary amino group linked to the reactive functionality; and (b) characterizing the molecule by mass spectrometry.

Abstract: Methods, reagents and kits are provided for the production and use in detection assays of labeled nucleic acid molecules wherein a labeling molecule is attached directly to the 3? end of the nucleic acid molecules.

Abstract: A system and method for forming encoded microparticles is described. One embodiment includes an encoded microparticle, the microparticle comprising a plurality of segments aligned along an axis, wherein the plurality of segments define a code for the microparticle; and an outer cuboid encapsulating the plurality of segments, wherein the plurality of segments are detectable through the outer cuboid.

Abstract: The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a poly-histidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6.

Abstract: The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a poly-histidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6.

Abstract: The present invention provides a powerful technique based on ultra high-throughput sequencing that finds structural aberrations of chromosomes and defines breakpoints. It is disclosed herein that, Anchored ChromPET, a technique to capture and interrogate targeted sequences in the genome, is a cost-effective means to identify chromosomal aberrations and define breakpoints. Using this method, we defined the BCR-ABL1 translocation DNA breakpoint to a base-pair resolution in Philadelphia chromosome positive cell lines and patient cells. This DNA-based method is highly sensitive and can detect signal using samples from which it is hard to obtain RNA or cells where the RNA expression has been silenced. These data demonstrate that ChromPET is a cost-effective and powerful technology that can identify and follow the appearance of chromosomal aberrations in various organisms, including, but not limited to, humans.

Abstract: The present invention provides methods and compositions for tagging nucleic acid sequence fragments, e.g., a set of nucleic acid sequence fragments from a single genome, with one or more unique members of a collection of oligonucleotide tags, or sequence tokens, which, in turn, can be identified using a variety of readout platforms. As a general rule, a given sequence token is used once and only once in any tag sequence. In addition, the present invention also provides methods for using the sequence tokens to efficiently determine variations in nucleotide sequences in the associated nucleic acid sequence fragments.

Abstract: The invention provides methods and compositions for attaching oligonucleotide tags to polynucleotides for the purpose of carrying out analytical assays in parallel and for decoding the oligonucleotide tags of polynucleotides selected in such assays. Words, or subunits, of oligonucleotide tags index submixtures in successively more complex sets of submixtures (referred to herein as “tiers” of submixtures) that a polynucleotide goes through while successive words are added to a growing tag. By identifying each word of an oligonucleotide tag, a series of submixtures is identified including the first submixture that contains only a single polynucleotide, thereby providing the identity of the selected polynucleotide. The analysis of the words of an oligonucleotide tag can be carried out in parallel, e.g.

Abstract: The present invention provides methods and systems for the capture and enrichment of target nucleic acids and analysis of the enriched target nucleic acids. In particular, the present invention provides for the enrichment of targeted sequences in a solution based format.

Abstract: A micro bead having a digitally coded structure that is partially transmissive and opaque to light. The pattern of transmitted light is determined by to decode the bead. The coded bead may be structured a series of alternating light transmissive and opaque sections, with relative positions, widths and spacing resembling a 1D or 2D bar code image. To decode the image, the alternating transmissive and opaque sections of the body are scanned in analogous fashion to bar code scanning. The coded bead may be coated or immobilized with a capture or probe to effect a desired bioassay. The coded bead may include a paramagnetic material. A bioanalysis system conducts high throughput bioanalysis using the coded bead, including a reaction detection zone and a decoding zone.

Abstract: A novel encoding system, compositions for use therein and methods for determining the source, location and/or identity of a particular item or component of interest is provided. In particular, the present invention utilizes a collection of one or more sizes of populations of semiconductor nanocrystals having characteristic spectral emissions, to “track” the source or location of an item of interest or to identify a particular item of interest. The semiconductor nanocrystals used in the inventive compositions can be selected to emit a desired wavelength to produce a characteristic spectral emission in narrow spectral widths, and with a symmetric, nearly Gaussian line shape, by changing the composition and size of the semiconductor nanocrystal. Additionally, the intensity of the emission at a particular characteristic wavelength can also be varied, thus enabling the use of binary or higher order encoding schemes.

Abstract: A method for detecting a target biological material using DNA barcodes is provided. The method is for detecting a target biological material (e.g., DNA) by using DNA barcodes by which a trace amount of target biological material can be detected in a rapid and economic manner without performing polymerase chain reaction (PCR). The method is characterized by the use of magnetic particles and polymer particles coated with DNA barcodes to sense a trace amount of a biological material (e.g., DNA).

Abstract: Methods for identifying strains of bacteria, particularly methods for serotyping Streptococcus pneumoniae in a sample, methods for detecting and/or classifying S. pneumoniae infection by serotype, array devices and kits for use in such methods are disclosed. Array devices comprise a set of capture antibodies immobilised on a substrate at pre-determined array positions, wherein the set of capture antibodies comprises serotype-distinguishing antibodies which differ in their binding specificity for different S. pneumoniae serotypes. Serotyping methods may employ whole cell detection utilising one or more detectable labels, including in situ labelling of array-bound cells.

Abstract: This optical coding device comprises a plurality of aggregates suitable for emitting infrared, visible or ultraviolet rays by luminescence, at least one of the said aggregates comprising at least one luminophore. This aggregate further comprises at least one particle consisting of a surface plasmon effect material, the said luminophore and the said particle being suitable for entering into interaction.

Abstract: A method for high throughput discovery of proteins fluorescently labeled at a cysteine residue and that undergo a change in fluorescence ratio at 2 wavelengths upon binding an unbound analyte is described. Probes are disclosed which are labeled at a cysteine residue and also probes labeled at both cysteine and lysine with two different fluorophores. These probes are useful for characterization and measurement of hydrophobic species in a fluid sample, particularly characterization and measurement of levels of unbound free fatty acids. A profile of unbound free fatty acids can be determined for an individual which can be used to determine the individual's relative risk for disease.

Abstract: Disclosed are a method for selective labeling of target nucleic acids on an array having nucleic acid analogue, e.g. PNA (peptide nucleic acid), probes immobilized on a support or supports, comprising adding to the array a detectable label and an agent for introducing the label into the target nucleic acids, after hybridization between the target nucleic acids and the nucleic acid analogue probes, and a method for detection of target nucleic acids using the same.

Abstract: A method of separating compounds, includes: a. tagging at least a first organic compound with a first tagging moiety to result in a first tagged compound; b. tagging at least a second organic compound with a second tagging moiety different from the first tagging moiety to result in a second tagged compound, the first tagging moiety and the second tagging moiety including at least one of a common repeat unit, but having a different number of the repeat units therein, the greater the number of repeat units, the greater the polarity of the tagging moiety; and c. separating the first tagged compound from a mixture including at least the second tagged compound using a chromatographic separation technique based upon differences in the number of repeat units between the first tagging moiety and the second tagging moiety.

Abstract: Families of compositions are provided as labels, referred to as eTag reporters for attaching to polymeric compounds and assaying based on release of the eTag reporters from the polymeric compound and separation and detection. For oligonucleotides, the eTag reporters are synthesized at the end of the oligonucleotide by using phosphite or phosphate chemistry, whereby mass-modifying regions, charge-modifying regions and detectable regions are added sequentially to produce the eTag labeled reporters. By using small building blocks and varying their combination large numbers of different eTag reporters can be readily produced attached to a binding compound specific for the target compound of interest for identification. Protocols are used that release the eTag reporter when the target compound is present in the sample.

Abstract: A novel tagged triazine library is provided in which three building blocks are prepared separately and assembled orthogonally to yield 1536 highly pure compounds. Each library compound contains a variety of a triethyleneglycol (TG) linker at one of the diversity sites of the triazine scaffold.

Abstract: The present invention relates to a process for synthesizing biopolymers by stepwise assembly from protected synthesis building blocks which carry two-stage protective groups. The two-stage protective groups are activated by a first illumination step and eliminated by a subsequent chemical treatment step. Photoactivatable components which considerably speed up the activation process via intramolecular triplet sensitizers or/and have fluorescence properties are used. The two-stage protective groups can be used in particular within the framework of quality control.

Abstract: Provided are recombinant pentapeptide repeat family proteins comprising at least one mutation of an i?1, i+1, and/or i+2 amino acid residue. Also provided are vectors comprising a nucleic acid sequence encoding the above-described pentapeptide repeat family proteins, protein libraries comprising the above pentapeptide repeat family proteins, and vector libraries comprising the above-described vectors. Additionally provided are methods of identifying a pentapeptide repeat family protein with an assayable phenotype, methods of labeling a nucleic acid-interacting macromolecule, and methods of detecting a nucleic acid-interacting macro-molecule.

Abstract: The invention provides compositions and methods for generating a molecular profile of genomic DNA by hybridization of labeled nucleic acid representing the genomic DNA to immobilized nucleic acid probes, e.g., arrays or biochips.

Abstract: Nature evolves biological molecules such as proteins through iterated rounds of diversification, selection, and amplification. The present invention provides methods, compositions, and systems for synthesizing, selecting, amplifying, and evolving non-natural molecules based on nucleic acid templates. The sequence of a nucleic acid template is used to direct the synthesis of non-natural molecules such as unnatural polymers and small molecules. Using this method combinatorial libraries of these molecules can be prepared and screened. Upon selection of a molecule, its encoding nucleic acid template may be amplified and/or evolved to yield the same molecule or related molecules for re-screening. The inventive methods and compositions of the present invention allow for the amplification and evolution of non-natural molecules in a manner analogous to the amplification of natural biopolymer such as polynucleotides and protein.

Abstract: The present invention provides a low cost imaged-based system for detecting, measuring and/or counting labeled features of biological samples, particularly blood specimens. In one aspect, the invention includes a system for imaging multiple features of a specimen that includes one or more light sources capable of successively generating illumination beams each having a distinct wavelength band and a plurality of differentially excitable labels capable of labeling a specimen comprising multiple features, such that each different feature is labeled with a different differentially excitable label.

Abstract: Alternatively spliced RNA, along with their normally-spliced counterparts, can be rapidly identified by hybridizing cDNA from normal tissue to cDNA from an abnormal or test tissue. The two cDNA populations are separately tagged prior to hybridization, which allows isolation of double-stranded cDNA containing both normal and alternatively spliced molecules. Within this population, pairing of cDNA molecules representing an alternatively spliced mRNA with cDNA molecules representing the counterpart normally spliced mRNA will form double-stranded cDNA with single-stranded mismatched regions. The mismatched double-stranded cDNA are isolated with reagents that bind single-stranded nucleic acids. The strands of each mismatched double-stranded cDNA are then coupled and analyzed, simultaneously identifying both normal and alternatively spliced molecules.

Abstract: A combinatorial library of solid-state fluorescent dyes is prepared by reacting an aldehyde with a solid-supported pyridinium salt having a linker.

Abstract: An assembly of a carrier having one or more reporter beads non-covalently attached thereto which may be used in relation to oligomer libraries. The oligomer libraries may be formed by a combinatorial split-process-recombine procedure. The oligomer library comprises a plurality of molecules comprising a multiplicity of different chemical groups. Each reporter bead has a different marker associated therewith to identify the chemical group attached to the carrier as well as to identify the position in sequence of the chemical group relative to other chemical groups in each molecule of the library. The markers are selected from fluorophores, chromophores, bar codes or radioactive or luminescent labels.

Abstract: A device and method for ablating tissue includes a sheath system with an occlusion structure. In certain embodiments, arrhythmias originating in pulmonary veins are treated cryogenically.