Abstract: The invention is directed to a Method for detecting a target moiety in a sample of biological specimens by: a) providing at least one conjugate with the general formula (I) An-P-Bm-Cq-Xo??(I) ?with A: antigen recognizing moiety; P: enzymatically degradable spacer; B: first binding moiety C second binding moiety X: detection moiety; n, m, q, o integers between 1 and 100, wherein B and C are non-covalently bound to each other and A and B are covalently bound to P b) labelling the target moiety recognized by the antigen recognizing moiety A with at least one conjugate c) detecting the labelled target moiety via detecting moiety X d) cleaving Cq-Xo by disrupting the non-covalent bond between Bm and Cq from the labelled target moiety e) cleaving the binding moiety Bm from the labelled target moiety by enzymatically degrading spacer P. The method is useful to identify target moieties on the biological specimens.

Abstract: The invention is directed to a detection reagent for CAR expressing cells according to general formula (1) Xn—Sm—Do wherein X is a polypeptide comprising at least 20 amino acids of which at least two amino acids are connected by at least one disulfide group S is a spacer comprising at least 15 carbon atoms D is a detection moiety m, o are integers independently between 1 and 200 n is an integer between 4 and 200 with the provisio that X binds to the antigen binding domain of a chimeric antigen receptor of a cell. The detection agent can be used in a method to detect cells having an antigen binding domain of a chimeric antigen receptor (CAR) binding the detection reagent and preferable to remove the detection moiety D from the detected cell.

Abstract: The present invention provides a method for analyzing simultaneously multiple human antigen-specific cell populations of a sample, the sample comprising B cells and antigen-specific cells, the method comprising a) separation of B cells from said sample, b) dividing the B cells into n sub-samples, c) differentially labeling the B cells of said sub-samples, wherein at least n-1sub-samples are labeled, d) pulsing of the B cells of each sub-sample with single or multiple peptides, e) pooling of the labeled and peptide-pulsed B cells with cells of said sample comprising said antigen-specific cells, f) co-cultivation of the cells of step e), g) flow cytometry analysis of the B cells with regard to their cell number and CD83 expression, thereby determining the potency of said antigen-specific cells in said sample.

Abstract: The present invention provides a process for generation of genetically modified T cells, T cell subsets and/or T cell progenitors comprising the steps: a) providing a cell sample comprising T cells, T cell subsets and/or T cell progenitors b) preparation of the cell sample by centrifugation c) magnetic separation of the T cells, T cell subsets and/or T cell progenitors d) activation of the enriched T cells, T cell subsets and/or T cell progenitors using modulatory agents e) genetic modification of the T cells, T cell subsets and/or T cell progenitors f) expansion of the genetically modified T cells, T cell subsets and/or T cell progenitors in a cultivation chamber g) washing of the cultured T cells, T cell subsets and/or T cell progenitors characterized in that all steps are performed in a closed and sterile cell culture system.

Abstract: The invention provides a system that comprises pharmaceutical agents for use in immunotherapy for reducing the side-effects of an antigen-recognizing receptor against antigen-expressing non-target cells in an individual. The system includes an antigen-recognizing receptor that specifically recognizes an antigen on target cells and at least on one hematopoietic cell type in the individual. The antigen-recognizing receptor is exemplified by chimeric antigen receptors (CAR) be expressed on the surface of an immune effector cells. The system also includes hematopoietic cells resistant to recognition of the same antigen by the antigen-recognizing receptor.

Abstract: The present invention provides a method for polyclonal stimulation of T cells, the method comprising contacting a population of T cells with a nanomatrix, the nanomatrix comprising a) a matrix of mobile polymer chains, and b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the T cells; thereby activating and inducing the T cells to proliferate; wherein the nanomatrix is 1 to 500 nm in size. At least one first and one second stimulatory agents are attached to the same or to separate mobile matrices. If the stimulatory agents are attached to separate nanomatrices, fine-tuning of nanomatrices for the stimulation of the T cells is possible. Closed cell culture systems also benefit from this method.

Abstract: The present invention provides a method for analyzing simultaneously multiple human antigen-specific cell populations of a sample, the sample comprising B cells and antigen-specific cells, the method comprising a) separation of B cells from said sample, b) dividing the B cells into n sub-samples, c) differentially labeling the B cells of said sub-samples, wherein at least n-1sub-samples are labeled, d) pulsing of the B cells of each sub-sample with single or multiple peptides, e) pooling of the labeled and peptide-pulsed B cells with cells of said sample comprising said antigen-specific cells, f) co-cultivation of the cells of step e), g) flow cytometry analysis of the B cells with regard to their cell number and CD83 expression, thereby determining the potency of said antigen-specific cells in said sample.

Abstract: The present invention provides a method polyclonal stimulation of T cells, the method comprising contacting a population of T cells with a nanomatrix, the nanomatrix comprising a) a flexible matrix, wherein said matrix is of polymeric material; and b) attached to said polymeric flexible matrix one or more polyclonal stimulatory agents which provide activation signals to the T cells; thereby activating and inducing the T cells to proliferate; wherein the nanomatrix is 1 to 500 nm in size. At least one first and one second stimulatory agents are attached to the same or to separate flexible matrices. If the stimulatory agents are attached to separate beads, fine-tuning of nanomatrices for the stimulation of the T cells is possible.

Abstract: The invention provides a method for convenient analysis and cell separation of antigen-specific T cells based on one or more products secreted by these cells in response to antigen stimulation. The T cells are provided with a capture moiety for the product, which can then be used directly as a label in some instances, or the bound product can be further labeled via label moieties that bind specifically to the product and that are labeled with traditional labeling materials such as fluorophores, radioactive isotopes, chromophores or magnetic particles. The labeled cells are then separated using standard cell sorting techniques based on these labels. Such techniques include flow cytometry, magnetic gradient separation, centrifugation, and the like.

Abstract: The invention provides a method for convenient analysis and cell separation of antigen-specific T cells based on one or more products secreted by these cells in response to antigen stimulation. The T cells are provided with a capture moiety for the product, which can then be used directly as a label in some instances, or the bound product can be further labeled via label moieties that bind specifically to the product and that are labeled with traditional labeling materials such as fluorophores, radioactive isotopes, chromophores or magnetic particles. The labeled cells are then separated using standard cell sorting techniques based on these labels. Such techniques include flow cytometry, magnetic gradient separation, centrifugation, and the like.

Abstract: A method is provided for modifying cells retained in a high gradient magnetic cell separation column (HGMS). Selected cells within a heterogeneous mixture are labeled with a label comprising a magnetic particle and specific for the selected cells. The labeled, selected cells are applied to the magnetic cell separation column, which retains the selected cells. The selected cells are then modified while retained in the column and then removed to the original suspension or to a purified, homogenous suspension.