Patent ID: 6265229
Filing Date: 2001-07-24
Classification: G01N,Y10S

Abstract:
A method for detecting a specific target cell in a cell suspension, the cell suspension comprising a mixed cell population, a fluid system containing a mixed cell population, or a homogenous cell population prepared from a solid tissue, without detection of normal and malignant hematopoietic cells, the method comprising:a. coating paramagnetic particles with an antibody or antibody fragment reactive with an antigen membrane structure specifically expressed on the target cell and not on a non-target cell in the cell suspension;b. contacting the coated paramagnetic particles with the cell suspension;c. incubating and rotating the mixture of coated paramagnetic particles and cell suspension;d. incubating the mixture of coated paramagnetic particles and cell suspension with an additional antibody or antibody fragment that is the same or different as that stated in (a), and binding the additional antibody or fragment to an antigen membrane structure specifically expressed on the target cell, that is the same or different as that stated in (a), and wherein the additional antibody or fragment is labeled;e. separating particle-target cell complexes from unbound particles, unspecifically bound non-target cells and unbound non-target cells in the mixture of coated paramagnetic particles and cell suspension by transferring the mixture to a separating apparatus, the separating apparatus comprising a filter having a pore size and shape capable of retaining particle-target cell complexes or rosettes and which filter provides a matrix for cell growth;f. growing cells of the separated particle-target cell complexes on the filter; andg. detecting labeled antibody/target cell/particle-immobilized antibody, labeled target cell/particle-immobilized antibody, or labeled antibody/target cell complexes and counting the complexes.