Preparation Of Rh-Leptin Monoclonal Antibody And Establishment Of The Method Of Detect Serum Leptin In Breast Cancer Patients

Objective We purified recombined human rh-leptin and prepared monoclonal antibody of recombined human leptin. We identified the biology characteristics of human rh-leptin and established the approach to detect the patient serum leptin levers of breast carcinoma. Methods The immunological activity of recombined human leptin (rh-leptin) after the protein renaturation and purification was identified in our research with SDS-PAGE. The mice were immunized with human rh-leptin, the MCAb were bolting by indirect enzyme-linked immunosorbant assay (ELISA), and the hybridoma cell's chromosome were analysed by colchicine; the subtype of the MCAb were identified with ISO-2 Mouse Monoclonal Antibody Isotyping Reagents, and the potency of the mcAb were detected. We established the approach to detect the patient serum leptin levers of breast carcinoma with indirect-ELISA and estimated the precision and accuracy of the approach. Results Two hybridoma cell lines named 6A5 and 3F7 were obtained. The cell lines stably secreted monoclonal antibody of recombined human leptin and subtype of the monoclonal antibody are IgG1 and IgG2b. We established the approach to detect the patient serum leptin levers of breast carcinoma with indirect-ELISA. Conclusion We purified recombined human rh-leptin and prepared monoclonal antibody of recombined human leptin. The approach to detect the patient serum leptin levers of breast carcinoma with indirect-ELISA could be used in detecting the patient serum leptin levers of breast carcinoma.