with every 100 ml of pbs n wash buffer composed of tween 20 0 5 ml diluting with every 100 ml of pbs o peg preparation solution composed of dmso 1 ml and 7 5 sodium bicarbonate 0 1 ml diluting with every 8 9 ml of incomplete 1640 cultivate solution 2 1 cell cultivation myeloma cells and hybridoma cells were cultured in complete 1640 cultivate solution and placed in 37 u00b0 c 5 carbon dioxide incubator changed liquid every other day and passaged once every 3 4 days 2 2 preparation of antigen synthesize a polypeptide with sequence of seq id no 1 and take the polypeptide as an antigen to immunize mice the specific preparation process of monoclonal antibody is conducted referring to u201c experimental technology of cellular and molecular immunology u201d chapter ii editor jin boquan fourth military medical university press 2001 11 isbn 7 81086 027 5 which is specifically described as follows balb c mice 4 8 weeks old female are immunized by the synthetic polypeptide seq id no 1 with specific steps as follows a first immunization polypeptide 500 u03bcg per mouse an equal volume of freund 39 s complete adjuvant is added and mixed followed by multi point injection subcutaneously with 1 ml per balb c mouse and 0 2 ml per point with an interval of 3 weeks b second immunization with the same dose and.