Dissertation
Dissertation > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Veterinary Pharmacology > Veterinary Toxicology > Chemical residues in animal foods

Establishment of Hybridoma Cell Lines Secreting Monoclonal Antibodies Against Sulfonamides

Author ZhangHaiLing
Tutor MaZuo
School Harbin Institute of Technology
Course Of Food Science
Keywords Sulfa drugs Hapten Diazotization Cell fusion Hybridoma
CLC S859.84
Type Master's thesis
Year 2009
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Sulfa drugs (SAS) is a class of chemicals widely used in livestock production synthetic antibacterial agents, which has a stable, easy to store as well as the side effects is low. However, excessive use of these drugs can cause allergic reactions, and may cause cancer. Sulfa drug residues in foods of animal origin caused by environmental pollution and human health hazards has been attracting much attention, so many countries sulfa drugs maximum residue levels (MRL) are clearly defined. For the sulfa drug residues in many ways, a variety of physical and chemical methods such as gas chromatography (GC) and mass spectrometry (MS), high performance liquid chromatography (HPLC). Although these methods are sensitive and accurate, but tedious sample pre-treatment step, to spend a long time, the high cost limits its widespread use. A quick and easy sulfa drug residue detection technology research and development is imminent. The enzyme-linked immunosorbent assay (ELISA) and GICA (GICA) fast, easy, specificity and high sensitivity, is an effective method of drug residue detection. The purpose of this study is a monoclonal antibody hybridoma lines secreting anti sulfa drugs, basic materials for the establishment of sulfa drugs immunological detection methods. This study used chemically synthesized sulfa drugs mother nucleus structure hapten, and by nuclear magnetic resonance apparatus and a liquid chromatography - mass spectrometry is used to identify the desired product. Diazotization method synthetic hapten and sulfa drugs (sulfaguanidine, Sulfathiazole, sulfadiazine, sulfamethazine, sulfamethoxazole oxazole) coupled to bovine serum albumin and ovalbumin identification successfully coupled with ultraviolet spectroscopy, infrared spectroscopy. Coupled fully immunized BALB / c female mice of 6 to 7 times, with the double agar diffusion method and indirect ELISA serum titer, when the indirect ELISA titer of 1:6400 or more, take the high titer mice spleen cells with SP2 / 0 myeloma cells to cell fusion with HAT medium, screening the culture, small colonies appeared in the fusion after the first 4 to 5 days, the 10th day hybridoma cells covered the hole bottom 60%, with the establishment of indirect ELISA screening positive wells screened positive hybridoma cells. This study successfully synthesized three kinds of hapten and the diazo coupling successfully prepared immunogen and coating antigen. The use of cell fusion was successfully prepared hybridoma lines secreting anti sulfa drugs, monoclonal antibodies, laid a solid foundation for the establishment of the immunological detection of sulfa drugs.

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