Application In Diagnosis Of Small Cell Lung Carcinoma And Preparation Of Recombinant Human Pro-gastrin Releasing Peptide And Its Monoclony Antibody

Lung cancer is the most common human malignant tumors, its morbidity and mortality is the most highest in all total tumors, World Health Organization Branch IARC reported the global incidence of lung cancer was1.608million in2008and was the first incidence about12.7%of all cancer. Small cell lung cancer is the highest malignancy in the subtypes of primary lung cancer and the worst prognosis by5-year survival rate of only1%-7%. Small cell lung cancer is about20%-25%in recently diagnosed lung cancer with more male patients; the ratio of male to female is2:1. it has transferred to extrapulmonary before being found because of its rapid growth, invasion and strong. If no effective treatment and intervention is carried out timely, the patient will die within a few months. Clinical early diagnosis of small cell lung cancer and the development of rational treatment methods and timely evaluation of the efficacy of treatment rate are particularly important for increasing the survival of the patient. ProGRP is the most valuable small-cell lung cancer serum tumor markers; the concentration of ProGRP31-98peptides in human serum is closely related to development and treatment of small cell lung cancer.For the establishment of human serum ProGRP31-98peptide early diagnosis detection kit for small cell lung cancer, we extracted the total RNA from small cell lung cancer tissue and designed primers for amplification ProGRP31-98peptide target gene according to the sequence of Genbank Nucleotide Sequence Database which was TA-cloned to pMD-18T vector, sequencing results showed that the sequence was consistent with the reported. The target gene of ProGRP31-98peptide was digested with BamH I and EcoR I from the T vector and cloned into the pGEM-2T expression vector with a GST purification tag and thrombin protease cleavage site which was transformed into bacterial BL21(DE3). The expression of ProGRP31-98fusion protein was induced by IPTG, the ProGRP31-98peptide was cut and purified from the fusion protein by thrombin. Further preparation of monoclonal antibodies and rabbit polyclonal antibodies were used to coating and marking, and ultimately establishing ProGRP31-98of the ELISA method for detection.We prepared antigens and antibodies for detection of small cell lung cancer By genetic engineering techniques and antibodies preparation at this research, finaly we established the ProGRP31-98detection EL1SA assay which will be a reliable method for the early diagnosis of small cell lung cancer, small cell lung cancer evaluation of the efficacy and improving the survival rate of small cell lung cancer patients.