Procaine monoclonal antibody hybridoma cell strain 4G6 and application thereof

Abstract

The invention discloses a procaine monoclonal antibody hybridoma cell strain 4G6 and application thereof, and belongs to the field of food safety immunodetection. A procaine complete antibody is completely mixed and emulsified with an equal amount of a Freund's adjuvant, and a BALB / c mouse is immunized by subcutaneous injection in the back of the mouse. A complete Freund's adjuvant is used for first immunization (80 mu g / mouse), an incomplete Freund's adjuvant is used for booster immunization of multiple times (50 mu g / mouse), and the procaine complete antibody (25 mu g / mouse, containing no adjuvant) is used for impact immunization for the last time. A high-tier and low-IC50 spleen cell of the mouse is extracted and fused with a myeloma cell of the mouse by virtue of a PEG (polyethylene glycol) method, and a hybridoma cell strain is obtained by screening and subcloning of three times with ic-ELISA (indirect competitive enzyme linked immunosorbent assay). A monoclonal antibody secretingby the cell strain has higher specificity and detection sensitivity (an IC50 value is 0.32ng / mL) to procaine, a raw material is provided for immunodetection of procaine residues in food, and high practical application value is achieved.

Description

technical field [0001] The invention relates to a procaine monoclonal antibody hybridoma cell line 4G6 and an application thereof, belonging to the technical field of food safety immunology detection. Background technique [0002] Procaine, also known as "Novocaine", is a white crystalline powder that is easily soluble in water. Its aqueous solution is acidic and unstable. It is easily hydrolyzed and oxidized by external conditions and turns yellow. It is often referred to as its salt. Salts are used as clinical local anesthetics. In addition, procaine penicillin, as the procaine salt of penicillin, is often used for moderate and mild infections caused by penicillin-sensitive Gram-positive cocci, and has good antibacterial activity. However, procaine penicillin is a kind of antibiotic drug, excessive use will bring great harm to human health, many countries have made strict restrictions on the content of procaine penicillin in animal food, especially in dairy products Among...

AI Technical Summary

Problems solved by technology

However, procaine penicillin is a kind of antibiotic drug, excessive use will bring great harm to human health, many countries have made strict restrictions on the content of procaine penicillin in animal food, especially in dairy products Among them, the Ministry of Agriculture of the People's Republic of China stipulated that the limit of procaine penicillin in milk should be ≤0.004mg / kg in the Announcement No. 235 on Veterinary Drug Residues

[0003] The determination of procaine content can be used as an important means to monitor the content of procaine penicillin, but at present, the determination of procaine penicillin mostly relies on large-scale instruments, such as gas chromatography, liquid chromatography, etc., and the cost is high , long time, high professionalism, difficult to achieve mass detection

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