Preparation and purification method of antibody for inhibiting chronic rejection of transplanted organ

Abstract

The invention discloses a preparation and purification method of an antibody for inhibiting a chronic rejection of a transplanted organ. The preparation and purification method comprises the following steps of: step 1, collecting a periphery blood mononuclear cell through a lymphocyte separating medium and carrying out induction culture with 20ng/ml MICA; step 2, extracting a mononuclear cell protein through a nuclear protein and cytoplasm protein extraction kit; step 3, carrying out series connection on a high performance liquid chromatography (HPLC) purification anti-MICA antibody; and step 4, identifying the anti-MICA antibody and using rMICA and WesternBlot to carry out a reverse authentication. The anti-MICA antibody prepared through the method can inhibit a genetic expression of an endogenous MICA by sealing an endogenous MICA protein, an endogenous MICA and an siRNA, and therefore, an expression effect of an MICA of a transplanted organ is blocked, namely, expressions of an MICA gene and an MICA protein in a donator organ after an organ transplantation operation are effectively reduced.

Description

technical field [0001] The invention relates to a preparation and purification method of an antibody for inhibiting chronic rejection of transplanted organs. Background technique [0002] Since the United States successfully implemented the world's first kidney transplant in the 1950s, organ transplantation, as an epoch-making new technology, has developed vigorously in the past 50 years, especially with the emergence of immunosuppressants in the late 1980s, It has greatly improved the success rate of organ transplantation and the survival rate of recipients. So far, more than 600,000 patients around the world have received organ transplant operations including kidney, liver, and heart. Since the implementation of organ transplantation in the 1960s, my country has ranked second in the world in the number of annual organ transplants by 2006, and has made considerable progress and development. Immunological rejection of grafts has been the biggest bottleneck that has plagued ...

AI Technical Summary

Problems solved by technology

At present, there are still some gaps in the research on MICA transplantation immunity at home and abroad: ① Under the condition of blocking the classic MHC molecules of the graft, some grafts still have acute and chronic rejection, indicating that there are other unknown factors that cause or participate in this disease. response, and MICA may be one of the more important factors

②Although anti-MICA antibodies have been found in some organs before and after kidney transplantation and in the lavage fluid of organs with immune rejection, the mechanism of transplant immune rejection involving MICA is still unclear, and most of the

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