MILTENYI BIOTEC B.V. & CO. KG

Method for determination of the kinetic binding parameters for a molecule binder conjugated to a fluorophore. Repeated staining of the antigen and detection of the emission radiation generates a titration series. Fitting of the emission radiation over the titration series allows for determination of kinetic binding parameters. If the emission radiation is detected with a camera, single pixel fitting using a global analysis permits discrimination of the specific binding signal from non-specific background in each pixel.

The present invention provides an in-vitro method for transferring one or more nucleic acids sequences comprising one or more transgenes into γδ T cells with a pseudotyped retroviral vector particle or a virus-like particle thereof, wherein said pseudotyped retroviral vector particle or virus-like particle thereof comprises a modified baboon endogenous retrovirus (BaEV) envelope glycoprotein, the method comprising the steps a) activation of γδ T cells, b) contacting said pseudotyped retroviral vector particle or virus-like particle thereof with said activated γδ T cells using a low concentration of said pseudotyped retroviral vector particle, c) expanding said genetically modified γδ T cells in the absence of an aminobisphosphonate and in the presence of IL-2 and IL-15, wherein said expansion is in the absence of feeder cells and in the absence of human serum, and wherein at least 75% of the transduced and expanded γδ T cells are CD45RA-γδ T cells.

The present invention provides a combination comprising a) an antigen binding domain specific for CD90, and b) an antigen binding domain specific for CD326, for use in treatment of human cancer comprising cancerous cells that co-express CD90 and CD326. In one embodiment of the invention the combination comprises a) an immune cell comprising a CAR comprising an antigen binding domain specific for a tag of a first and a second polypeptide, b) said tagged first polypeptide that has an antigen binding domain specific for CD90, and c) said tagged second polypeptide that has an antigen binding domain specific for CD326, wherein the tag of the first polypeptide and the tag of the second polypeptide are identical. In a further embodiment the concentrations used for said first and that second polypeptide are below the activation threshold of said CAR, respectively, but the sum of both concentrations is above the activation threshold of said CAR.

This disclosure provides a composition comprising i) a pseudotyped retroviral vector particle or virus-like particle thereof comprising a) one envelope protein with antigen-binding activity, wherein said envelope protein is a recombinant protein that does not interact with at least one of its native receptor(s) and is fused at its ectodomain to a polypeptide comprising an antigen binding domain specific for a tag of a tagged polypeptide, and wherein said envelope protein is protein G, HN or H derived from the Paramyxoviridae family, and b) one envelope protein with fusion activity derived from the Paramyxoviridae family, and ii) said tagged polypeptide, wherein said tagged polypeptide binds specifically to an antigen expressed on the surface of a target cell, thereby transducing the target cell with said retroviral vector particle or thereby inducing uptake of the virus-like particle into the target cell. A pharmaceutical composition thereof and an in vitro method for transduction of targets cells with said vector particle are also disclosed.

This disclosure provides a method and kit for analyzing compositions of human floorplate mesencephalic dopaminergic (mesDA) progenitor cells that are obtained, for example, by differentiating embryonic stem cells. The mesDA progenitors are characterized using the positive markers FOXA2. OTX2, and optionally NKX2. 1. in combination with the negative markers OCT3/4, and optionally PAX6, and SOXI. Cells with an appropriate protein expression profile have therapeutic potential for restoring the function of dopaminergic neurons in conditions such as Parkinson's Disease.

The invention is directed to a much faster way of combining multiple colour channels in fluorescence microscopy for complex fast diagnostic and research purposes. The advantage is that the procedure and the microscope setting can be simplified and miniaturised to an extend that the construction of such automatic systems become much easier and by omitting the erasing step the speed of analysis is faster and moved to the processing of the images by image processing systems, nowadays working at almost real time.

The invention is directed to a process for extracting target cells from a three dimensional biological specimen by the steps imaging the three dimensional specimen; identifying target cells; registering the spatial parameters (x,y,z coordinates) of the target cells; and extraction of target cells according to their spatial parameters

The invention provides a method which allows the separation of different workflow steps for barcoding of target nucleic acids and therefore providing optimal reaction conditions for each workflow step, especially for template switching reactions. Moreover this method provides the opportunity to perform the reactions such as barcoding reactions of two different nucleic acid molecules from one cell such as RNA and genomic DNA molecules in a single step. The method comprises the steps: (a) Providing a plurality of cells comprising target RNA molecules and at least one solid support comprising capture oligonucleotides for said target RNA molecules and barcode oligonucleotides; (b) Partitioning said plurality of cells and said solid supports such that each cell is included into a separate partition and each partition comprises a solid support; (c) Lysing said cell, thereby obtaining a mixture of target and non, target RNA molecules; (d) Hybridizing said target RNA molecules to the capture oligonucleotides for said target RNA molecules, thereby obtaining target RNA molecules attached to said solid support (e) Disrupting the partitions and separating the non-target RNA molecules from the target RNA molecules attached to said solid support (f) Generating double stranded nucleic acids from the target RNA molecules by nucleic acid synthesis, wherein the capture oligonucleotides serve as primer and the target RNA molecules serve as templates (g) Attaching the barcode oligonucleotides to the double stranded nucleic acids from target RNA molecules, thereby generating barcoded nucleic acids from target RNA molecules; Characterized in that in step a) said plurality of cells additionally comprise target genomic DNA molecules and said at least one solid support additionally comprise capture oligonucleotides for target genomic DNA molecules and in that the capture oligonucleotides for target RNA and target genomic DNA molecules are different.

An assembly ( 10 ) contains a aseptic flow of a liquid out of a container ( 20 ). The container ( 20 ) holding the liquid and an adapter ( 30 ) is configured to mechanically engage with the container ( 20 )

The invention relates to a system, comprising: a) a sample processing unit, comprising an input port and an output port coupled to a rotating container having at least one sample chamber, the sample processing unit configured provide a first processing step to a sample or to rotate the container so as to apply a centrifugal force to a sample deposited in the chamber and separate at least a first component and a second component of the deposited sample; and b) a sample separation unit coupled to the output port of the sample processing unit, the cell separation unit comprising separation column holder, a pump and a plurality of valves configured to at least partially control fluid flow through a fluid circuitry and a separation column positioned in the holder, the separation column configured to separate labeled and unlabeled components of sample flowed through the column.