title: Development of a Sensitive Luciferase-Based Sandwich ELISA System for the Detection of Human Extracellular Matrix 1 Protein.
abstract: Enzyme-linked immunosorbent assay (ELISA) has been one of the main methods for detecting an antigen in an aqueous sample for more than four decades. Nowadays, one of the biggest concerns for ELISA is still how to improve the sensitivity of the assay, and the luciferase-luciferin reaction system has been noticed as a new detection method with high sensitivity. In this study, a luciferin-luciferase reaction system was used as the detection method for a sandwich ELISA system. It was shown that this new system led to an increase in the detection sensitivity of at least two times when compared with the traditional horseradish peroxidase (HRP) detection method. Lastly, the serum levels of the human extracellular matrix 1 protein of breast cancer patients were determined by the new system, which were overall similar to the HRP chemiluminescent system. Furthermore, this new luciferase reporter can be implemented into other ELISA systems for the purpose of increasing the assay sensitivity.
authors.affiliation: 1 Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University , Xi'an, P.R. China .
authors.forename: Ya
authors.initials: Y
authors.lastname: Li
authors.affiliation: 1 Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University , Xi'an, P.R. China .
authors.forename: Yanqing
authors.initials: Y
authors.lastname: Li
authors.affiliation: 1 Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University , Xi'an, P.R. China .
authors.forename: Junli
authors.initials: J
authors.lastname: Zhao
authors.affiliation: 1 Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University , Xi'an, P.R. China .
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Zheng
authors.affiliation: 2 Department of Pathology, Northwestern University Feinberg School of Medicine Chicago , Chicago, Illinois.
authors.forename: Qinwen
authors.initials: Q
authors.lastname: Mao
authors.affiliation: 1 Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University , Xi'an, P.R. China .
authors.forename: Haibin
authors.initials: H
authors.lastname: Xia
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D001360:Avidin / Q000737:chemistry; D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D005260:Female; D049454:Firefly Luciferin / Q000737:chemistry; D015870:Gene Expression; D017930:Genes, Reporter; D057809:HEK293 Cells; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008156:Luciferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D049408:Luminescent Agents / Q000737:chemistry; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; C490607:ECM1 protein, human; D016326:Extracellular Matrix Proteins; D049408:Luminescent Agents; D011994:Recombinant Proteins; D001360:Avidin; D049454:Firefly Luciferin; D006735:Horseradish Peroxidase; D008156:Luciferases
keywords: extracellular matrix 1; horseradish peroxidase; luciferase; monoclonal antibody; sandwich ELISA