Rapid Preparation Of Monoclonal Antibodies By Intrasplenic Immunization And Treatment Of Colon Cancer By Tumor Vaccine Combined With Chemotherapeutics

1. Rapid preparation of monoclonal antibody(mAb) usingMuc1-Histag as antigen by intrasplenic immunizationSubcutaneous and muscular injections were generally taken as common ways ofimmunization to prepare antibodies. Instead, intrasplenic immunization, though withmore complex procedures, could raise antibody levels in mice in a much shorter time.Intrasplenic immunization was generally thought as an excellent option for boostingimmunization.Preparation of mAbs goes through multiple steps including immunization,hybridization, monoclonal hybrids selection, antibody production and different testsafterward. In this article, single intrasplenic injection was used to innoculateMucin(MUC)1-Histag into mice. As the results shown in the article, threemonoclonal hybrid cell lines were finally selected after series of protocols includingsingle intrasplenic immunization, two to three turns of monoclonal enlargements andselections. Indirect ELISA tests indicate that all three mAbs target at Histag ofMUC1-histag.Despite the result that all the mAb target at Histag of MUC1-histag, it is stillpossible that MUC1could be the target in the case that MUC1-Histag was theantigen. MUC1is a type of colon cancer associated protein and Histag is used as akey component in chromatography. Rapid preparation of mAb either against MUC1or Histag contributes a lot to the basic science and clinic study. Despite thesuccessful preparation of monoclonal antibodies using single time intrasplenicimmunization with shorten time period, many uncertain factors or potential problems possibly affect the final results during the process, such as construction andmolecular weight of antigen, concentration and lipopolysaccharide(LPS) level ofantigen solution and so on. Furthermore, comparing to the subcutaneousimmunization, less lymphocytes could be activated by single time intrasplenicimmunization, which directly narrows the types of monoclonal antibodiessubsequently. All these factors or problem are still needed to be further revealed.2． Treatment of colon cancer with different combinations of tumorvaccine,chemotherapeutics and CpG ODNIn China, colon cancer ranks third in all malignant cancer. One of the commonside effects of surgical treatments is the recurrence lead by the scattered tumor cells.In the experiments shown in the article, mice colon cancer models were made toimitate the recurrence that was designed to be treated with different combinations oftumor tissue lysates(TTL), capecitabine and CpG ODN. Especially the trial toactivate local immune cells by giving CpG ODN first, followed by TTL orcapecitabine or both potentially offer new theories or methods to either scientificresearches or clinic works.Subcutaneous tumors were made by back subcutaneous injections of coloncancer cells cultured in vitro to prepare TTLs. Mice colon cancer models were madeby intraperitoneally injections of minor amount of colon cancer cells. The psychicaland bodyweight changes of mice during the whole process after being givendifferent combined treatments were observed. The death date of each mouse wasrecorded to calculate survive rates. The results indicate different survive rates andbodyweight changes between groups under different combining therapies. Besides,mice treated with TTL combined with Capecitabin showed temporary losing of longmustaches.The results indicate that CpG ODN didn’t work efficiently as adjuvant in risingup survival rate of tumor-hearing mice. Capecitabin probably contributes to theprolonged survival differences between groups. Besides the groups designed in the article, other combinations and dosages of each component could still be tried toclarify the best option. During the treatment process, mice in the group treated withTTL combined with capetabine were observed to lose long moustaches, mechanismsof which need to to be further discussed and studied.