Preprint Article Version 1 This version is not peer-reviewed

Sea Bass Immunization to Downsized Betanodavirus Protein Displayed in The Surface of Dna-Damaged Repair-Less Bacteria

Version 1 : Received: 24 July 2019 / Approved: 26 July 2019 / Online: 26 July 2019 (11:46:15 CEST)

A peer-reviewed article of this Preprint also exists.

Lama, R.; Pereiro, P.; Novoa, B.; Coll, J. Sea Bass Immunization to Downsize the Betanodavirus Protein Displayed in the Surface of Inactivated Repair-Less Bacteria. Vaccines 2019, 7, 94. Lama, R.; Pereiro, P.; Novoa, B.; Coll, J. Sea Bass Immunization to Downsize the Betanodavirus Protein Displayed in the Surface of Inactivated Repair-Less Bacteria. Vaccines 2019, 7, 94.

Journal reference: Vaccines 2019, 7, 94
DOI: 10.3390/vaccines7030094

Abstract

This work describes practical immunization of European sea bass (Dicentrarchus labrax) juveniles against viral nervous necrosis virus (VNNV), a betanodavirus causing worldwide mortalities in many fish species. Protection was obtained with the so called spinycterin vehicles consisting in irreversibly DNA-damaged DNA-repair-less E.coli displaying at their surface a downsized antigen. In this work we, i) maximized bacterial expression levels by downsizing the C protein to a fragment (frgC91-220) containing most of its antigenicity, ii) developed an scalable autoinduction bacterial media based in soy-bean increasing membrane display and reproducibility, iii) enriched surface expression by screening different anchors from several prokaryotic origins (anchor+frgC91-220), iv) preserved frgC91-220 antigenicity by inactivating bacteria by irreversible DNA-damage by means of Ciprofloxacin, and v) increased safety using a repair-less E.coli strain as spinycterin chassis. These second generation of spinycterins protected fish against VNNV challenge with partial (Nmistic+frgC91-220) or 100 % (YBEL+frgC91-220 ) protection, in contrast to those fish immunized with frgC91-220 spinycterins. The proposed spinycterin platform has high levels of environmental safety and cost effectiveness, thus providing potential for small fish vaccines for sustainable aquaculture.

Subject Areas

VNNV; mass-immunization; sea bass; recombinant bacterins; spinycterins; DNA-damaged; repair-less

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