title: An Ultra-High Fluorescence Enhancement and High Throughput Assay for Revealing Expression and Internalization of Chemokine Receptor CXCR4.
abstract: Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high-throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip-coating poly(styrene-co-N-isopropylacrylamide) spheres on the glass substrate. The self- assembled spheres form three-dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF-1α activation in various cancer cells, even for those with extremely low expression level.
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: He
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Jun
authors.initials: J
authors.lastname: Lao
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Baosheng
authors.initials: B
authors.lastname: Ge
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Ren
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China.
authors.forename: Naseer Ullah
authors.initials: NU
authors.lastname: Khan
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, P. R. China. fhuang@upc.edu.cn.
authors.forename: Fang
authors.initials: F
authors.lastname: Huang
mesh_terms: D000178:Acrylamides / Q000737:chemistry*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D054377:Chemokine CXCL12 / Q000737:chemistry* / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry*; D005453:Fluorescence; D006801:Humans; D019718:Receptors, CXCR4 / Q000737:chemistry*; D015398:Signal Transduction; D013343:Styrenes / Q000737:chemistry*
chemical_list: D000178:Acrylamides; D054377:Chemokine CXCL12; D019718:Receptors, CXCR4; D013343:Styrenes; C000610368:poly(styrene-co-N-isopropylacrylamide)
keywords: CXCR4 expression internalization; fluorescence enhancement; high throughput; photonic crystals