| Melamine is an important organic chemical raw material. Because of its high nitrogen content and some other chemical properties, it is misused in feed and food based on the technical loopHoles in determination of doping protein,which has resultted in issues such as pet food contamination in 2007 and infant formula contamination in 2008. Previous studies show that animals which have take the melamine long-term would caused damage of the genitourinary system and stones in the bladder and kidney, and may further induced the bladder cancer and even death of them. After the incident of milk contamination, the melamine is also detected in candy, fried fritters, eggs and other foods.It can be found that people has access to melamine in daily life easily.The existing methods of detecting melamine are mainly traditional chemical methods such as turbidity method, the weight method and potentiometric titration and chromatograpHy methods such as GC-MS, HPLC, HC-MC and so on. The traditional chemical methods only to be applied for determination the purity of the melamine in chemical industry,and these methods cannot used to detect the traces of melamine in food. While the chromatograpHy methods need the good laboratory, specialized operators, expensive analytical instruments, complex testing procedures and the money for inspection cost, so these methods are not suitable for the rapid detection of large numbers of samples, and much less suitable for general family self-test use. To counter for this situation, the methods of detection melamine which principle based on immunological is developed, such as ELISA kit. One-time bulk test samples are the greatest features of ELISA kit which different from the above detection methods. However, the methods of ELISA kit still failed to shake off the shackles of equipment, and it need some basic technical personnel to operate, high cost, long time and its specificity is not high. So the high specificity and low cost methods of rapid detection melamine which suitable for on-site real-time of detection need to be developed.This research was based on immunological theory, immunized BALB/C mice with full antigen of melamine, then made the homologous mouse myeloma cells to fuse with the spleen cells of mice immunized. We used ELISA to screen the positive fusion hole and limited dilution to subcloning,and this is the basic method for hybridoma screening.After the 3-5 subcloning in the screening,we developed three monoclonal antibodies against melamine which named as 2A7,6A6 and 6E3.We also have done the karyotype analysis of the positive monoclonal hybridoma cells and the characterization of the three monoclonal antibodies.Then we made the monoclonal antibodies against melamine to be labelled by the colloidal nano-selenium particles which were preparated eligible. Finally the test strip labelled by selenium for rapidly detection melamine was assembled and installed, and we have done a preliminary test with it.The test strip which we have developed preliminary would make foundation for the rapid test of melamine, and it will be a simple, low cost and short time powerful tool which would applied for the rapid screening of melamine in the incident scene and self-test by family. |
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