Preparation method of regenerated gene protein REG1 alpha monoclonal antibody

Abstract

The present invention relates to a preparation method of a regenerated gene protein REG1 alpha monoclonal antibody. The method comprises the following steps: (1) preparing immunized mice; (2) culturing myeloma cells for 3-4 days to obtain cultured mouse myeloma cells; (3) dissolving REG1 alpha protein, inoculating abdominal cavity of the immunized mice with the dissolved REG1 alpha protein for immunization, obtaining splenocytes, and dispersing the splenocytes to obtain a splenocyte precipitate; (4) centrifuging the cultured mouse myeloma cells to obtain a myeloma cell precipitate; (5) respectively suspending the splenocyte precipitate and the myeloma cell precipitate by using a DMEM high-glucose culture solution, conducting mixing, and centrifuging the mixture to obtain a precipitate; (6)centrifuging and suspending the precipitate to obtain fusion cells; (7) culturing and determining the fusion cells, and determining hybridoma cells; (8) screening and separating monoclonal antibodies; and (9) preparing a large amount of monoclonal antibodies, namely producing the monoclonal antibodies from a hybridoma cell line according to a conventional method. The preparation method is rapid and high in specificity.

Description

technical field [0001] The invention relates to a preparation method of a monoclonal antibody, in particular to a preparation method of a regenerative gene protein REG1α monoclonal antibody. Background technique [0002] Abnormal expression of the regeneration family gene (Reg) family is closely related to various human diseases such as inflammation and tumors, especially digestive system diseases. Clinically, abnormally increased Reg protein can often be detected in pathological specimens and serum of tumor patients, so Reg protein can be used as a clinical marker for predicting tumor progression and progression, especially for digestive system tumors, and can also be used as a diagnostic marker and therapeutic target genes. Reg1 is a member of the type I subclass of the Reg gene family. Fukui et al. found that the expression of Reg1 in gastric cancer tissues can reach 37.3%. Reg1 positive patients have higher proliferating cell nuclear antigen index than negative patients...

AI Technical Summary

Problems solved by technology

At this stage, most of the antibodies for REG1α protein detection on the market are rabbit anti-polyclonal antibodies, which have relatively low sensitivity and specificity, while early clinical diagnostic samples are mainly serum and urine, which have a high sensitivity and specificity for detection. Higher requirements, therefore, a high titer monoclonal antibody against REG1α protein is the best way to improve the performance of the detection reagent