title: A novel bispecific antibody targeting tumor necrosis factor α and ED-B fibronectin effectively inhibits the progression of established collagen-induce arthritis.
abstract: Specific delivery of TNF-α antagonist to the inflamed site can increase its efficacy and reduce the side effects. In this study, we constructed a bispecific diabody (BsDb) that targets TNF-α and ED-B-containing fibronectin (B-FN), a fibronectin isoform specifically expressed in the pannus of the inflamed joint in rheumatoid arthritis. BsDb was produced in Escherichia coli as inclusion bodies, purified to homogeneity, and refolded to the functional form. Our data demonstrate that BsDb could simultaneously bind to human TNF-α and B-FN and neutralize TNF-α action. In the collagen-induced arthritis mouse model, we compared the biodistrubtion and therapeutic efficacy of BsDb with those of the anti-TNF-α scFv (TNF-scFv). Similar to TNF-scFv, BsDb penetrated into the synovial tissue quickly, showing a rapid clearance from blood and normal organs. In contrast, BsDb could selectively accumulate and retain in arthritic joints of mice for a long period time, resulting in a much stronger inhibition of arthritis progression in mice than TNF-scFv. The findings described herein indicate that BsDb has a good specificity to the inflamed joint, with low toxicity to normal organs and seems to be an ideal biological agent for the treatment of RA and other chronic inflammatory disease.
authors.affiliation: Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, 430062 Wuhan, China; Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, 430062 Wuhan, China. Electronic address: liumengy@tom.com.
authors.forename: Mengyuan
authors.initials: M
authors.lastname: Liu
authors.affiliation: Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, 430062 Wuhan, China; Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, 430062 Wuhan, China.
authors.forename: Mian
authors.initials: M
authors.lastname: Xie
authors.affiliation: Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, 430062 Wuhan, China; Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, 430062 Wuhan, China.
authors.forename: Sijing
authors.initials: S
authors.lastname: Jiang
authors.affiliation: College of Animal Sciences, Yangtze University, Jingmi Road 88-2, 434025 Jingzhou, China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Liu
authors.affiliation: Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, 430062 Wuhan, China; Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, 430062 Wuhan, China.
authors.forename: Lujun
authors.initials: L
authors.lastname: Li
authors.affiliation: Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, 430062 Wuhan, China; Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, 430062 Wuhan, China.
authors.forename: Dongxu
authors.initials: D
authors.lastname: Liu
authors.affiliation: Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, 430062 Wuhan, China; Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, 430062 Wuhan, China.
authors.forename: Xiaosong
authors.initials: X
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics / Q000627:therapeutic use*; D001169:Arthritis, Experimental / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D001483:Base Sequence; D045744:Cell Line, Tumor; D016207:Cytokines / Q000032:analysis / Q000378:metabolism; D018450:Disease Progression; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics / Q000627:therapeutic use*; D005528:Foot / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics / Q000627:therapeutic use; D014018:Tissue Distribution; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics / Q000627:therapeutic use*
chemical_list: D018033:Antibodies, Bispecific; D016207:Cytokines; C067625:FN1 protein, human; D005353:Fibronectins; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha
keywords: Bispecific antibody; ED-B-containing fibronectin; Pharmacokinetics; Rheumatoid arthritis; Tumor necrosis factor α (TNF-α)