title: Single domain antibody multimers confer protection against rabies infection.
abstract: Post-exposure prophylactic (PEP) neutralizing antibodies against Rabies are the most effective way to prevent infection-related fatality. The outer envelope glycoprotein of the Rabies virus (RABV) is the most significant surface antigen for generating virus-neutralizing antibodies. The small size and uncompromised functional specificity of single domain antibodies (sdAbs) can be exploited in the fields of experimental therapeutic applications for infectious diseases through formatting flexibilities to increase their avidity towards target antigens. In this study, we used phage display technique to select and identify sdAbs that were specific for the RABV glycoprotein from a naïve llama-derived antibody library. To increase their neutralizing potencies, the sdAbs were fused with a coiled-coil peptide derived from the human cartilage oligomeric matrix protein (COMP48) to form homogenous pentavalent multimers, known as combodies. Compared to monovalent sdAbs, the combodies, namely 26424 and 26434, exhibited high avidity and were able to neutralize 85-fold higher input of RABV (CVS-11 strain) pseudotypes in vitro, as a result of multimerization, while retaining their specificities for target antigen. 26424 and 26434 were capable of neutralizing CVS-11 pseudotypes in vitro by 90-95% as compared to human rabies immunoglobulin (HRIG), currently used for PEP in Rabies. The multimeric sdAbs were also demonstrated to be partially protective for mice that were infected with lethal doses of rabies virus in vivo. The results demonstrate that the combodies could be valuable tools in understanding viral mechanisms, diagnosis and possible anti-viral candidate for RABV infection.
authors: CAS Key Laboratory for Pathogenic Microbiology and Immunology (CASPMI), Institute of Microbiology, Chinese Academy of Sciences, Beijing, China ; University of Chinese Academy of Sciences, Beijing, China.
authors: Bhargavi M
authors: BM
authors: Boruah
authors: Dawei
authors: D
authors: Liu
authors: Duan
authors: D
authors: Ye
authors: Tie-Jun
authors: TJ
authors: Gu
authors: Chun-Lai
authors: CL
authors: Jiang
authors: Mingsheng
authors: M
authors: Qu
authors: Edward
authors: E
authors: Wright
authors: Wei
authors: W
authors: Wang
authors: Wen
authors: W
authors: He
authors: Changzhen
authors: C
authors: Liu
authors: Bin
authors: B
authors: Gao
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000145:classification / Q000235:genetics / Q000276:immunology*; D000914:Antibodies, Viral / Q000145:classification / Q000235:genetics / Q000276:immunology*; D002161:Camelids, New World; D064236:Cartilage Oligomeric Matrix Protein / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D051379:Mice; D019151:Peptide Library; D010802:Phylogeny; D055503:Protein Multimerization; D011818:Rabies / Q000276:immunology / Q000517:prevention & control*; D011819:Rabies Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D011820:Rabies virus / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D061905:Single-Domain Antibodies / Q000145:classification / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D064236:Cartilage Oligomeric Matrix Protein; D019151:Peptide Library; D011819:Rabies Vaccines; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; D014759:Viral Envelope Proteins; C577553:TSP5 protein, human
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