The Application Of Cim-Chip In Leucocythemia Immuno-typing

Cell Immuno-Chip (Cim-Chip) is a kind of Bio-Chip which could be used to detect the cell surface antigens by directly inspecting the cells with a microscope on the basis of protein chip and the specific reaction between the cell surface antigens and the associated antibodies. Cim-Chip has the advantage of high throughput and could be used to detect up to several hundreds of cell surface antigens simultaneously. In this paper, the experimental conditions of Cim-Chip were optimized. The technique was used exploringly on the immnuophenotyping of leukemia and the detection of drug curative effect of cancer and the approving result was obtained. The major contributions include the following aspects:1. The fabricating conditions of the Cim-Chip were evaluated. CD45 monoclonal antibody was immobilized on a chip. After the reaction between the immobilized antibody and the leukocyte solution, the capture efficiencies of leukocytes were compared among the different conditions such as different incubating times and different methods of dyeing. The optimized experimental conditions established the experimental fundament for the investigation and appliance of the Cim-Chip.2. The application exploration of the Cim-Chip on the immuophenotyping of leukemia. Monoclonal antibodies such as CD2,CD3,CD5,CD10,CD22,CD30,CD33,CD45,CD117 were used as the markers for immuophenotyping of leukemia, The common antibody of leukocyte CD45 was used as positive control for the test. In our work, 4 kinds of leukocytes were incubated with Cim-Chip, the surface antigens expressed by each kind of the leukocyte were observed to validate the utilization feasibility of the Cim-Chip on the immunophenotyping of leukemia. The Cim-Chip is also reacted with different samples from patients, the images of the results were captured and compared by the results of cytometry. Convincing conclusion was reached that different kinds of surface antigens expressed by different types of leukemia.3. The exploration of the Cim-Chip on the drug efficiency and drug screening. Different drugs were added into culture medium respectively during the cell culture, and the leukocytes were reacted with the Cim-Chip to observe the combination between the leukocytes and the monoclonal antibodies. The changes of the cell surface antigen expression were evaluated and the anticancer drugs were screened by curative...