title: A Versatile Microarray Immobilization Strategy Based on a Biorthogonal Reaction Between Tetrazine and Trans-Cyclooctene.
abstract: Given its increasing importance in transforming biomedical research in recent years, microarray technology has become highly popular as a powerful screening platform in detecting biomolecule interactions, discovering new inhibitors, and identifying biomarkers as well as diagnosing disease. The success of microarray technology in various biological applications is highly dependent on the accessibility, the functionality, and the density of the surface bound biomolecules. Therefore, compound immobilization represents a critical step for the successful implementation of microarray screening. Herein we describe a fast and site-specific microarray immobilization approach by using trans-cyclooctene-tetrazine ligation. This approach not only ensures fast immobilization and uniform display of biomolecules, but also allows the optimum orientation of biomolecules after immobilization. All these excellent properties facilitate subsequent interactions of the biomolecules and their interacting partners during the screening process. We envision that the immobilization strategy described here can find useful applications in many other microarray related studies.
authors.affiliation: Department of Biology and Chemistry, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong, China.|Key Laboratory of Biochip Technology, Biotech and Health Centre, 15 Shenzhen Research Institute of City University of Hong Kong, Shenzhen, 518057, People's Republic of China.
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, Singapore, 138669, Singapore.
authors.forename: Liqian
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Biology and Chemistry, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong, China.|Key Laboratory of Biochip Technology, Biotech and Health Centre, 15 Shenzhen Research Institute of City University of Hong Kong, Shenzhen, 518057, People's Republic of China.
authors.forename: Haipeng
authors.initials: H
authors.lastname: Lei
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, Singapore, 138669, Singapore. sslee@ibn.a-star.edu.sg.
authors.forename: Su Seong
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Department of Chemistry, National University of Singapore, Singapore, Singapore.
authors.forename: Shao Q
authors.initials: SQ
authors.lastname: Yao
authors.affiliation: Department of Biology and Chemistry, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong, China. hongysun@cityu.edu.hk.|Key Laboratory of Biochip Technology, Biotech and Health Centre, 15 Shenzhen Research Institute of City University of Hong Kong, Shenzhen, 518057, People's Republic of China. hongysun@cityu.edu.hk.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Sun
mesh_terms: D000595:Amino Acid Sequence; D019817:Benzoic Acid / Q000737:chemistry; D034242:Cyclooctanes / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D046228:Microarray Analysis / Q000379:methods*; D010455:Peptides / Q000737:chemistry
chemical_list: D034242:Cyclooctanes; D006573:Heterocyclic Compounds, 1-Ring; D010455:Peptides; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019817:Benzoic Acid
keywords: Fast kinetic; Microarray; Site-specific immobilization; TCO–tetrazine ligation