Preparation Of Monoclonal Antibodies Against Listeria Monocytogens

In recent years, microbiological contamination caused by food-borne diseases and potential health hazards has become the major public health problem of the world. At present, in our country, the examination method of the pathogenic bacteria mainly to use traditional the GB code method, but its examination time is too long and the sensitivity is not high, so it is disadvantageous to the food-borne disease prevention and the control, poses the serious threat to consumer's health of our country. WHO has listed the Listeria monocytogenes(LM) as one of the four big food pathogenic bacteria of the 1990s (enteropathogenic E.coli, Clostridium botulinum, Aeromonas hydrophila and LM), simultaneously it has been listed as one of the twelve kinds of pathogenic microorganisms which is the 21st century to have to the Chinese health the significant influence , the disease lethality caused by it has reached as high as 30~40%. In order to improve the examination method of this fungus, we have developed the antiuninuclear cell proliferation Liszt bacillus monoclonal antibody.This research uses the inactivate monocaryotic proliferation Lisztthe bacillus entire fungus immune female BALB/c mouse, when the blood valence at the mouse tail vein achieve above1: 10000, takes its spleen and carries on the cytomixis according to the conventional method with the SP2/0 myeloma cell , the proportion of the spleen cell and the SP2/0 myeloma cell is 8: 1, carries on screening by the indirect ELISA method, then clone to the strong masculine list clone hole,after five limited dilution method clones, obtains four hybridoma cell to be able stably to secrete the monoclonal antibody ,and named them 1B9, 2C3, 4D8 and 4H6, appraisal to its subgroup, respectively is IgG1, IgG1, IgG2b and IgG1;the difference of the Four generation valence is not big, that shows secretion immune body ability is quite stable; The chromosome appraisal, respectively is 104, 106, 103, 107 chromosomes, conforms to the hybridoma cell's characteristic; Paring experiment demonstrate that ,1B9 and 2C3, 4D8 and 1B9, 4D8 and2C3, 4H6 and 1B9, 4H6 and the 2C3 monoclonal antibody separately aims at the different antigen epi-position, 4H6 and 4D8 monoclonal antibody are view of same antigen epi-position; The overlapping response demonstration, only has 4D8 and the backwoods coli has the slight overlapping response.