Development and optimization of an antibody array method for potential cancer biomarker detection
Shuangshuang Wang
Ping Zhao
Brian Cao
摘要：Biomarkers play an important role in the detection at an early stage of pancreatic cancer.The aim of the present study was to optimize the conditions of antibody arrays for detecting Hippocalcin-like 1(HPCAL1),phosphati-dylethanolamine binding protein 1(PEBP1),lectin galactoside-binding soluble 7(LGALS7),and serpin peptidase inhibitor clade E member 2(SERPINE2)as biomarkers for pancreatic cancer detection in a single assay and to investigate antibodies' specificity and cross-reactivity.Capture antibodies against HPCAL1,PEBP1,LGALS7and SERPINE2 were printed on nitrocellulose coated glass slides.HPCAL1,PEBP1,LGALS7 and SERPINE2proteins with different concentrations were incubated with the capture antibodies at different temperatures for different time periods.Biotinylated detection antibodies recognizing a different epitope on the captured proteins and a secondary detection molecule(Streptavidin-PE)were used to detect fluorescent signals.The arrays showed the strongest signals when the concentration of the capture antibodies was at 500 μg/mL in PBST0.05(PBS with0.05% Tween-20),and the slides were incubated overnight at 4℃.The lowest protein concentration for detection was 2 ng/mL.Each antibody demonstrated high specificity to the corresponding antigen in detecting a mixture of4 proteins without significant cross-reactivity.The fluorescence and biomarker concentration displayed a linear correlation.The antibody microarray system could be a useful tool for potential biomarker detection for pancreatic Cancer.
机标关键词：detectionmicroarraypancreatic cancerdifferentbinding proteinfluorescencesignificantTween-20
分类号：R3(基础医学)
资助基金：This work was partially supported by the Lustgarten Foundation for Pancreatic Cancer Research(RFA-08-003)
论文发表日期：2011-01-01
在线出版日期：2025-08-15（本平台首次上网日期，不代表文献的发表时间）
英文信息