title: [Preparation of monoclonal antibodies against enterovirus type 71 with an epitope-incorporated adenovirus type 3 vector].
abstract: Objective To develop the monoclonal antibodies (mAbs) against enterovirus type 71 (EV71). Methods Two neutralization epitopes, SP70 and SP55, from EV71 were cloned into the hexon gene of adenovirus type 3 to generate a recombinant adenovirus type 3 (R1R2A3) presenting SP70 and SP55 antigens. BALB/c mice were immunized with the R1R2A3. The mAbs were developed with hybridoma technology and were analyzed with microneutralizing assay, indirect ELISA, Western blotting and direct immunofluorescence assay (DFA). Results The study obtained four hybridoma cell clones, 2C4, D2C9, I2G2 and I12C3. ELISA showed that the titer of D2C9 against EV71 was 1:8 000 000 and the titers of 2C4, I2G2, and I12C3 all were 1:500 000. ELISA and Western blotting demonstrated that all mAbs could specifically recognize the VP1 of EV71. In addition, D2C9 recognized the SP70 epitope, and 2C4, I12C3 and I2G2 all recognized the SP55 epitope. DFA revealed that all mAbs could react with EV71, but not with Coxsackie virus A16 (CoxA16). Conclusion Four mAbs against EV71 have been developed successfully, and all of them could react with EV71 rather than CoxA16.
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510230, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510230, China.
authors.forename: Xingui
authors.initials: X
authors.lastname: Tian
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510230, China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Xue
authors.affiliation: Guangzhou RIDA Biotech Company, Guangzhou 510000, China.
authors.forename: Minglong
authors.initials: M
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510230, China.
authors.forename: Zhichao
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510230, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Li
authors.affiliation: Guangzhou RIDA Biotech Company, Guangzhou 510000, China. *Corresponding authors, E-mail: zhourong@vip.163.com.
authors.forename: Chenyang
authors.initials: C
authors.lastname: Li
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510230, China. *Corresponding authors, E-mail: biolcy@126.com.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhou
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D029821:Enterovirus A, Human / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008807:Mice, Inbred BALB C; D017386:Sequence Homology, Amino Acid; D014709:Vero Cells
chemical_list: D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D000939:Epitopes; C018315:hexon capsid protein, Adenovirus