title: Site-specific labeling of baculovirus in an integrated microfluidic device.
abstract: Labeling of viruses can be used to reveal viral infection pathways and screen potential anti-viral drugs. Complex procedures, including virus cultivation, purification and labeling are involved in traditional virus labeling methods. And the manipulation of living virus brings risk to researcher health. In this work, we report a general method for site-specific labeling of the envelope virus in an integrated microfluidic device with simple procedures and high security. Site-specific labeling of virus was achieved by fusing the biotin acceptor peptide (AP-tag) and the biotin ligase enzyme (BirA enzyme) with the envelope protein GP64 of baculovirus. The AP-tag could be modified by BirA enzyme to introduce the biotin moiety onto the viral envelope. Western blots and fluorescence colocalization analysis proved that the baculoviruses were biotinylated and labeled with high efficiency. The integrated device incorporated several operation steps including cell seeding, cell culture, cell transfection, virus culture and virus labeling. Since virus biotinylation was achieved during the process of virus cultivation, the complex procedures of virus labeling were simplified in our device. Furthermore the whole process could be completed in the integrated microfluidic device, and direct contact between viruses and researchers could be eliminated in our method, which could greatly reduce the risk to researcher health during living virus labeling.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, P R China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Shu
authors.forename: Wen
authors.initials: W
authors.lastname: Lu
authors.forename: Shu-Lin
authors.initials: SL
authors.lastname: Liu
authors.forename: Na
authors.initials: N
authors.lastname: Xu
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhen-Hua
authors.initials: ZH
authors.lastname: Zheng
authors.forename: Dai-Wen
authors.initials: DW
authors.lastname: Pang
authors.forename: Han-Zhong
authors.initials: HZ
authors.lastname: Wang
authors.forename: Zhi-Ling
authors.initials: ZL
authors.lastname: Zhang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D015153:Blotting, Western; D002232:Carbocyanines / Q000737:chemistry; D019731:Carbon-Nitrogen Ligases / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D061987:Sf9 Cells; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D002232:Carbocyanines; D029968:Escherichia coli Proteins; D006023:Glycoproteins; D010455:Peptides; D012097:Repressor Proteins; D014759:Viral Envelope Proteins; C089187:cyanine dye 3; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D001710:Biotin; D019809:Streptavidin; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli