Prokaryotic Expression And Production Of Monoclonal Antibodies To Fpr3

FKBP is a protein family existing widely in biological system.There are four members of FKBP family that have been found in the yeast:Fpr1, Fpr2, Fpr3, Fpr3 is nucleolar proteins, which involves in ribosomes assembling, leading ribosomes or protein concerned to cross the nuclear pore complex Fpr3 also can regulate cell cycle. Fpr3 can could inhibit cell cycle progression in the G2 when the abnormality of cell cycle emerges.Because of the importance of Fpr3 physiological function, we hope to analyse its space structure, despite the fact that it is difficult to crystallize. Monoclonal antibody is a kind of antibody that is specially for antigen epitopes.when single protein can not crystallize,it,however, can crystallize with its antibody.The binding of antibody and its protein can stabilize the conformation of protein, which can improve the probability of protein crystallization. Here antibody act as a molecular chaperones that can bind with unstable protein,assisting the protein to fold,assemble,transport and degradate by controlling the binding and ilivery, not act as a ingredient when protein domains perform functions.This study transformed the plasmid pEGX-5X-1-fpr3 into E.coliBL21, induced by IPTG to express GST-Fpr3 fusion protein, and then GST-Fpr3 fusion protein was purifiedwith which mousse were immunized. Two cell lines, which secrete monoclonal antibody of Fpr3 protein stably after cell fusion, were obtain. The molecular weight of the antibody is 152kDa.The subclass of antibody is IgGl, the antibody titer is 1.5×104, 1.0×105 respectively. Also identified by Western blot to the specificity of monoclonal antibodies, the final results show that two hybridomas secreting monoclonal antibodies can recognize Fpr3 protein Specifically. The binding specificity of antigen and its antibody play a very important role in the study of structure and function of Fpe3 protein.