Establishment Of Hybridoma Cell Lines Secreting Monoclonal Antibody Against Enrofloxacin

Enrofloxacin (Enro) is a specific medicine for animals, which belongs to Fluoroquinolone .it is a new type man-made synthetic broad-spectrum sterilizing antibiotic. At present, it has been widely used in veterinary and fish. And it has already become one of frequently used drugs in veterinary and fish in our country. However, accumulative residue of Enro in food produce can cause severe problems because of its illegal use, and gives highly rise to concern regarding consumer safety. Normal bacterium in human will get drug-resistance if people eat contaminated food. Also, it causes intoxication, hypersensitivity. Synthesis of nuclear acid will be disturbed, and people are prone to cancers.Many countries, such as America, European United, Japan and China included, have formulated strict regulations for the use of Enro. Enro has been prohibited using in USA.And in addition, it has approved by European Community that the Maximum Residue limits (MRL) of Enro in edible animal tissue are not beyond 30ug/kg. The residue of Enro is a focal point in exports and imports quarantine. An effective and quick detect technology is required to fulfill the quarantine system to ensure regular exports and imports trades. Also, it will restrict the illegal use of Enro, insure food safety, and improve people healthy.Many chemical and physics methods for detecting Enrofloxacin residued in edible tissues of animals have been established. Though some of these methods are sensitive and accurate, they are not suitable for screening a large number of samples because these methods are time-consumption, costly and highly skilled. Enzyme-linked immunosorbent ?ssay (ELISA) as a rapid, special and sensitive biological method, is gradually applied in drug residues assay, It is a new technology combinated antigen-antibody special reaction with sensitive enzyme reaction, which is highly sensitive, selective, quick and convenient. The immune analysis replies to microanalysis from complicated samples.Monoclonal antibodies are produced by cell fusion technology. In contrast to polyclonal antibodies, monoclonal antibodies have following virtues: identical structures, uniform component, and high specificity. Animate biotic-activity and fewer cross reacts.First, Enro was coupled with carrier protein bovine serum album (BSA) and ovalbumin (OVA) to prepare the immune antigen Enro-BSA and the detection antigen Enro-OVA by N-hydroxvsuccinimide ester (NHS) method respectively. Conjugated antigens were analysized by the UV scanning and animal experiment. The results showed that the conjugated antigens were successfully obtained and could be used to prepare the monoclonal antibody against Enro with immune antigen Enro-BSA and detection...