Preparation Of Anti-Her2 Full Human Monoclonal Antibody And Applacation Of Breast Cancer Cells Fluorescence Labeling

Single molecular tumor fluorescence imaging technique mainly aim at the specific target of tumor cells, and obtain a targeting specific tumor cell immune fluorescent marker by connecting highly specific antibody with no biotoxic Fluorescent quantum dots.This method can detect cancer timely and determine the effectiveness of the target.Single molecule tumor cell imaging has become a hot research topic after the molecular target drug.Human epidermal growth factor receptor 2(Her2) is a member of the epidermal growth factor receptor family, study prove that Her2 can affect cell growth,development and differentiation, It also play a key role in the case of amplification and conversion about several kinds of tumor cells.Especially in breast cancer, more than a third of breast cancer patients over expressed Her2.so that Her2 is an important target to diagnosis and identification of breast cancer.Currently, anti-Her2 monoclonal antibodies has been developed in the field of diagnosis and treatment for breast cancer. but in clinical,tumor imaging study also use mouse antibody or humanized antibody, this kind of antibody in the body exist the risk of human anti mouse antibody response(HAMA).Useing the technology of antibody engineering to develop high affinity full human monoclonal antibody will solve this problem.This study selected Her2 as a target. After four round of screening,we obtained a high antigen-affinity monoclonal phage by using fixed Her2 antigen in a large phage display Fab library.Expressed Fab and Ig G1 peotein,ELISA analysis found that these two antibodies can specific bind to Her2 protein.SPR analysis indicated that two antibodies with high affinity of target Her2(Fab=15n M,Ig G1=1.01 n M).Flow cytometry also proved that antibodies can specific bind to SK-BR3 cell lines(Her2+),but no combination with MCF-7(Her2-).We labeled anti-Her2 Ig G1 with azido and produced a immune fluorescent marker(Her2-Pdots)that can specific target breast cancer cell line(SK-BR3) by connecting anti-Her2 Ig G1-N3 with a newtype of high stability fluorescent polymer quantum dots(labeled DBCO),and the study of single molecule imaging to SK-BR3 cell line by confocal microscope found that breast cancer cell line can be recogined and fluorescence imaging by Her2-Pdots.As a result,we screened and expressed a full human anti-Her2 antibody with high affinty,and this antibody can be used for SK-BR3 cell line specific imaging by connecting a new type of polymer quantum dots. Achieving the function of Her2-labeling and breast cancer cells imaging.