Thesis (Ph.D.) - Cairo University - Faculty of Agriculture - Department of Agriculture Biochemistry

Recently, the spread of epidemics has increased in conjunction with the ability of microbes to develop themselves and increase their ability to resist antimicrobial, which forced scientists to search for unconventional methods of treatment and quick and sensitive methods of diagnosis.In this research the study was done using camel to produce different Nano-antibodies those have the ability to diagnose and treat different types of pathogenic bacteria (Mannheimiahaemolytica, Salmonellatyphimurium, Salmonellaarizonae, and Klebsiellapneumoniae), which characterized by cost, skills and production requirements are lower than the traditional antibodies.The present work developed a sandwich ELISA-based Nano- antibody towards Mannheimia haemolytica,Salmonella typhimurium, Salmonella arizonae, and Klebsiella pneumoniae as the first report for the treatment of them. VHH amplified fragments at 450 bp were isolated and cloned into the phagemid vector pMECS, then via phage display technique isolate the specific Nano-antibody against each strain, then screening by ELISA to obtain the top 5 AD value Nano- antibody for each strain. Nano-antibodies protein was purified under non-denaturing conditions. The purifiedNano-antibodies were chractraized and used for Competed ELISA, it is show binding to the specific strain, and also, there was a cross-reaction with the other strains. Nano-antibodies are expected to survive in harsh conditions, such as those found in the stomach gut, creating opportunities for nano-antibodies' oral delivery to treat different diseases as gastrointestinal diseases. But these Nano- antibodies cannot use in diagnostic. So, many trials should be done to improve its sensitivity in the future, and can use the epitops that characterizes each strain as antigen to immunizing the camel to produce the nano antibody used for diagnostic

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