Cost effective filamentous phage based immunization nanoparticles displaying a full-length hepatitis B virus surface antigen

Abstract

Hepatitis B virus (HBV) is one of the major causes of chronic hepatitis, cirrhosis and liver cancer. In combating HBV infections, HBV diagnosis and vaccination are therefore critical. The hepatitis B virus surface antigen (HBsAg) is a key target molecule in developing vaccines and diagnostic systems. To date, although HBsAg has been expressed in bacteria, yeasts and mammalian cells, there are still limitations in the existing ones, which leave the necessity for searching new HBsAg production methods. In this study, a simple phage display-based method was developed to produce the purified full-length HBsAg molecules for further immunization studies. For this purpose, the HBsAg coding gene was cloned into a pCANTAB5E phagemid vector and expressed on the surface of M13 filamentous phages. The HBsAg-expressing phage nanosystem was then used as immunization agent in BALB/cJ mice. The ELISA results for sera obtained from mice immunized with HBsAg-displaying phage particles revealed an immune response against HBsAg. These results demonstrate the potential use of a full-length antigen to be displayed on phages as cost effective adjuvant-free immunization agents as an alternative to the highly purified and more expensive antigens conjugated with carrier molecules.

A novel phage-displayed MilA ELISA for detection of antibodies against Myc. bovis in bovine milk

“…It makes the recombinant proteins more expensive, time-consuming and hard to make in uniform batches (Du & Rehm, 2017). The large-scale production of the phage-displayed antigens is less expensive, more uniform and easier than other methods (Baclioglu et al, 2014). Additionally, phage-displayed antigens are more resistant to tough environmental conditions and keep their uniformity for longer time compared to the bacterially expressed recombinant proteins (Aghebati-Maleki et al, 2016).…”

“…Ligation product was transfected to electrocompetent E. coli TG1 (Agilent) cells using the electroporation method described by Green and Sambrook (Green & Sambrook, 2012). The transformed colonies were screened for the presence of milA ‐pCANTABF12 construct by colony PCR using vector‐specific primers (Baclioglu et al, 2014) and sequencing.…”

Beyond phage display: non-traditional applications of the filamentous bacteriophage as a vaccine carrier, therapeutic biologic, and bioconjugation scaffold

“…Initially, the major advantages to phage display of such antigens were speed, ease of purification and low cost of production ( Gram et al, 1993 ). E. coli F17a-G adhesin ( Van Gerven et al, 2008 ), hepatitis B core antigen ( Bahadir et al, 2011 ), and hepatitis B surface antigen ( Balcioglu et al, 2014 ) all elicited antibody responses when displayed on pIII, although none of these studies compared the immunogenicity of the phage-displayed proteins with that of the purified protein alone. Phage displaying Schistosoma mansoni glutathione S -transferase on pIII elicited an antibody response that was both higher in titer and of different isotypes compared to immunization with the protein alone ( Rao et al, 2003 ).…”

Cloning of anti-hbsag single chain variable fragments from hybridoma cells for one-step elisa

“…The Lig7 anti-HBsAg scFv was amplified using 2 oligonucleotides containing NdeI and NotI restriction enzyme recognition sites (underlined): TUB ç 721 ç NdeI (5 0 -TCGCCATATGCAGGCCCAGGTG-CAGCTGCAGGAGTCAGG-3 0 ) and Lig ç 7 ç Reverse ç Not ç I (5 0 -GAGTCATTCTGCGGCCGCCCGTTTGATTTCCAGCT TGG-3 0 ) and digested with the appropriate restriction enzymes. The digested product was then inserted into the NdeI/NotIdigested pQE2/AP vector [26].…”