Anti-glycocholic acid monoclonal antibody and preparation method thereof

Abstract

The invention discloses an anti-glycocholic acid monoclonal antibody and a preparation method thereof. In the present invention, high-concentration cholic acid and glycine small molecules are added to the enzyme-labeled antibody. If the antibody can be combined with the small molecule to occupy the binding site of the antibody, so that it will no longer bind to the enzyme-labeled antibody, the ELISA test result is negative. , Excluding antibodies that can only recognize cholic acid and glycine, while reducing the clone screening of positive cell lines, ascites preparation, purification, etc., greatly increasing the probability of obtaining monoclonal antibody cell lines that only recognize glycocholic acid. The method of the invention effectively expands the screening range under the premise of reducing the workload, so that the probability of obtaining the desired monoclonal antibody cell line is improved. Through the established screening method, the present invention obtains a monoclonal antibody that can specifically recognize glycocholic acid, which solves the problems of large differences among batches of polyclonal antibodies, poor reproducibility, and high cost. At the same time, it also reduces the cross-reaction with other substances, and can be used for the competitive detection of ELISA and chemiluminescent method of glycocholic acid.

Description

technical field [0001] The invention relates to a monoclonal antibody and a preparation method thereof, in particular to an anti-glycocholic acid monoclonal antibody and a preparation method thereof, and the invention belongs to the field of biotechnology. Background technique [0002] Cholylic acid (Cholyglycine, CG) is a conjugated cholic acid formed by the combination of cholic acid and glycine. It is one of the main cholic acids. The normal metabolic pathway of CG is entero-hepatic circulation, which is mainly synthesized in liver cells, that is, cholesterol is converted into primary bile acids through complex enzymatic reactions, including cholic acid (CA) and chenodeoxycholic acid (CD). -CA), drains into the gallbladder through the capillary and bile ducts, enters the duodenum and small intestine along with the bile, and participates in the digestion and absorption of fat. [0003] Under normal circumstances, 95% of bile acids are reabsorbed in the terminal ileum, and...

AI Technical Summary

Problems solved by technology

This method has a heavy workload. In order to increase the probability of screening CG monoclonal antibodies and reduce the workload, the present invention establishes a detection method for the preparation of CG monoclonal antibodies that is different from the traditional method: through one cell fusion experiment, using different carrier CG-coupled antigen is coated on the microtiter plate, and then added to the cell culture supernatant according to the conventional indirect ELISA, and then the enzyme-labeled antibody mixture containing high concentrations of bile acid and glycine small molecules (0.1mg / ml) is added for competition inhibition test , screen positive cell lines, and obtain cell lines that have no cross-reaction with bile acid and glycine

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