Abstract: The present invention provides a nucleotide sequence for encoding CAR, a ROBO1 CAR-NK cell of expressing the CAR, and preparation and application thereof. The ROBO1 CAR-NK cell provided by the present invention can specifically kill tumor cells by using ROBO1 antibody for the construction of CAR-NK cells and using ROBO1 molecules as target antigens. It can be used as a therapeutic agent for tumor diseases, for the treatment of tumor with highly expressing of ROBO1, without harmful phenomena such as cytokine release syndrome, thus providing new treatments for the tumors which are ineffective in traditional surgery, chemotherapy and radiotherapy. It has lower toxicity, higher safety and better specific lysis activity compared with ROBO1 CAR-T cell.

Abstract: The present invention relates to chimeric antigen receptor cells targeting ROBO1, in particular, enhanced CAR-T cells and CAR-NK cells targeting ROBO1, and preparation and application thereof. The cells can stably expressing CAR elements, while secreting extracellular domain molecules expressing PD-1 protein or mutants thereof, and thus may block PD-11PD-L1 molecular interaction. It has been found through animal experiments that the cells have very good anti-tumor effects, and the above-mentioned cells can significantly reduce tumor recurrence and improve the survival rate compared with the conventional ROBO1-targeted CAR modified cells.

Abstract: Disclosed are a bispecific antigen-binding construct and the preparation method and use thereof, wherein the construct comprises a first antigen binding unit and a second antigen binding unit, the first antigen binding unit is a single chain variable region antibody fragment ScFV which specifically binds to the surface antigen of immune cells, and the second antigen binding unit is a Slit2D2 protein fragment which specifically binds to the surface antigen Robo1 of tumour cells. That is to say, the construct can bind to the surface antigen of immune cells and the surface Robo1 molecule of tumour cells at the same time, so that as the distance between tumour cells and immune cells get smaller, the quiescent immune cells are effectively activated, and the effect of killing and wounding tumours is produced.

Abstract: Provided is a method for modifying a chimeric antigen receptor-modified T cell (CAR-T cell). The method comprises expressing an ScFv-CD8-4-1 BB-CD3? molecule in a T cell.

Abstract: Disclosed are a bispecific antigen-binding construct and the preparation method and use thereof, wherein the construct comprises a first antigen binding unit and a second antigen binding unit, the first antigen binding unit is a single chain variable region antibody fragment ScFV which specifically binds to the surface antigen of immune cells, and the second antigen binding unit is a Slit2D2 protein fragment which specifically binds to the surface antigen Robo1 of tumour cells. That is to say, the construct can bind to the surface antigen of immune cells and the surface Robo1 molecule of tumour cells at the same time, so that as the distance between tumour cells and immune cells get smaller, the quiescent immune cells are effectively activated, and the effect of killing and wounding tumours is produced.

Abstract: Provided is a method for modifying a chimeric antigen receptor-modified T cell (CAR-T cell). The method comprises expressing an SCFV-CDS TM-4-1BB-CD3? molecule in a T cell. The CAR-T cell prepared using the method can specifically recognize and bind to a tumor cell with elevated expression of a ROBO1 protein, and can be used to prevent and treat a corresponding tumor-related disease.

Abstract: Disclosed are a bispecific antigen-binding construct and the preparation method and use thereof, wherein the construct comprises a first antigen binding unit and a second antigen binding unit, the first antigen binding unit is a single chain variable region antibody fragment ScFV which specifically binds to the surface antigen of immune cells, and the second antigen binding unit is a Slit2D2 protein fragment which specifically binds to the surface antigen Robo1 of tumour cells. That is to say, the construct can bind to the surface antigen of immune cells and the surface Robo1 molecule of tumour cells at the same time, so that as the distance between tumour cells and immune cells get smaller, the quiescent immune cells are effectively activated, and the effect of killing and wounding tumours is produced.

Abstract: Provided are a PD-1 CAR NK-92 cell and a preparation method and use thereof. The PD-1 CAR NK-92 cell expresses PD-1-CD8™-4-1BB-CD3? fusion protein in NK-92 cells. The PD-1 CAR NK-92 is obtained by infecting an NK92 cell line with a PD-1 CAR molecule and obtaining monoclonal cells by means of flow screening, and culturing and expanding CAR NK92 monoclonal cell strains with stable traits and a high killing activity. The cells can be produced on a large scale, can be used in different patients without GVHR rejection, and have a specific killing activity and significant therapeutic effect on tumors.

Abstract: A chimeric antigen receptor (CAR) and a gene encoding the CAR. The CAR comprises an extracellular domain capable of binding to an antigen, a transmembrane domain, and intracellular immune co-stimulatory molecule, wherein the extracellular domain comprises a D2 domain of a Slit2 protein. A chimeric antibody-expressing cell, which introduces a gene encoding the CAR into a cell so as to express the CAR on the surface of the cell. The CAR or CAR-expressing cell can be used as a cell drug for the treatment of tumor diseases. By using the CAR for engineering cells, especially T cells, the engineered T cells can specifically recognize and kill tumors, and have higher tumoricidal activity.

Abstract: The present invention provides a recombinant fusion protein. The fusion protein is formed by the fusion of D2 domain of Slit2 protein and HSA protein, and the position 386 amino acid of the Slit2 protein molecule is serine.

Abstract: Disclosed are a bispecific antigen-binding construct and the preparation method and use thereof, wherein the construct comprises a first antigen binding unit and a second antigen binding unit, the first antigen binding unit is a single chain variable region antibody fragment ScFV which specifically binds to the surface antigen of immune cells, and the second antigen binding unit is a Slit2D2 protein fragment which specifically binds to the surface antigen Robo1 of tumour cells. That is to say, the construct can bind to the surface antigen of immune cells and the surface Robo1 molecule of tumour cells at the same time, so that as the distance between tumour cells and immune cells get smaller, the quiescent immune cells are effectively activated, and the effect of killing and wounding tumours is produced.

Abstract: Disclosed is a fusion protein Slit2D2-HSA formed by fusion of a Slit2D2 polypeptide and a human serum albumin HSA, and use thereof for the manufacture of a medicament for prophylaxis and/or treatment of sepsis, and the said fusion protein Slit2D2-HSA retains the pharmacological activities of Slit2 in inhibition of neutrophil migration and treatment of sepsis, and has an improved stability, prolonged half-life and improved therapeutic effect of sepsis. Compared with protein of Slit2, the polypeptide of Slit2D2 has a smaller molecular weight which is much easier to be purified and separated in preparation and used in the development of drugs.

Abstract: Provided is a method for modifying a chimeric antigen receptor-modified T cell (CAR-T cell). The method comprises expressing an SCFV-CDS TM-4-1BB-CD3? molecule in a T cell. The CAR-T cell prepared using the method can specifically recognize and bind to a tumor cell with elevated expression of a ROBO1 protein, and can be used to prevent and treat a corresponding tumor-related disease.

Abstract: Provided are a PD-1 CAR-T cell, a preparation method thereof, and an application thereof. By means of chimeric antigen receptor-modified T cell transformation, PD-1-CD8™-4-1BB-CD3? molecules are expressed in a T cell. The CAR-T cell prepared by the method can specifically recognize and bind with tumor cells with high PDL-1 protein expression, and is applicable to preparation of a drug for preventing and treating tumor diseases.