Aggregation and antigenicity of virus like particle in salt solution—A case study with hepatitis B surface antigen

Chen, Yi; Zhang, Yan; Quan, Can; Luo, Jian; Yang, Yanli; Yu, Mengran; Kong, Yuanyuan; Ma, Guanghui; Su, Zhiguo

doi:10.1016/j.vaccine.2015.03.078

Cited by 30 publications

(23 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For instance, traditional chromatography typically utilizes low ionic strength buffers for sample binding and high ionic strength buffer for elution. AF4 has been successfully used for assaying the quality and quantity of viruses and VLPs (Chen et al 2015;Chuan et al 2008;Eskelin et al 2016;Lang et al 2009;Somasundaram et al 2016;Wei et al 2007). However, in most cases purified specimens have been used.…”

Section: Discussionmentioning

confidence: 99%

“…AF4 has been widely used for separation and characterisation of submicron-and micron-sized bioparticles including viruses [for reviews see (Ratanathanawongs and Williams 2006;Roda et al 2009)]. It has been exploited to optimise virus-like particle (VLP) production in bacterial, yeast and mammalian cells, to assay VLP assembly and disassembly conditions, to determine size distribution of sample components in virus and VLP specimens, to quantitate particle amounts, and to study the presence of aggregates and fragments in particle preparations (Chen et al 2015;Chuan et al 2008;Lang et al 2009;Somasundaram et al 2016;Wei et al 2007). The majority of AF4 applications on viruses and VLPs have used specimens purified to near homogeneity.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

et al. 2017

View full text Add to dashboard Cite

Viruses come in various shapes and sizes, and a number of viruses originate from extremities, e.g. high salinity or elevated temperature. One challenge for studying extreme viruses is to find efficient purification conditions where viruses maintain their infectivity. Asymmetrical flow field-flow fractionation (AF4) is a gentle native chromatography-like technique for size-based separation. It does not have solid stationary phase and the mobile phase composition is readily adjustable according to the sample needs. Due to the high separation power of specimens up to 50 µm, AF4 is suitable for virus purification. Here, we applied AF4 for extremophilic viruses representing four morphotypes: lemon-shaped, tailed and tailless icosahedral, as well as pleomorphic enveloped. AF4 was applied to input samples of different purity: crude supernatants of infected cultures, polyethylene glycol-precipitated viruses and viruses purified by ultracentrifugation. All four virus morphotypes were successfully purified by AF4. AF4 purification of culture supernatants or polyethylene glycol-precipitated viruses yielded high recoveries, and the purities were comparable to those obtained by the multistep ultracentrifugation purification methods. In addition, we also demonstrate that AF4 is a rapid monitoring tool for virus production in slowly growing host cells living in extreme conditions.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

et al. 2017

View full text Add to dashboard Cite

show abstract

“…Also, the use of ammonium sulfate (AS) during purification, and the presence of salt in the purified samples may result in formation of aggregated particles. Recently, it has been shown that the presence of salt can lead to aggregation of HBsAg particles into oligomeric particles [17]. …”

Section: Resultsmentioning

confidence: 99%

“…Structural characterization of hepatitis B VLPs formed by HBsAg oligomerization has been undertaken by various researchers and shows that the particles display an octahedral symmetry [14] and that significant structural changes occur upon heat treatment [15]. It has also been shown that processing steps following production, purification, and reassembly of HBsAg into VLPs can affect the structural properties of the vaccine produced in yeast [16, 17]. In the oral maize delivery system, no purification is required, but removal of lipids is a preferred step to produce a highly thermostable vaccine [10].…”

Section: Introductionmentioning

confidence: 99%

Biochemical and biophysical characterization of maize-derived HBsAg for the development of an oral vaccine

Shah

Hayden

Fischer

et al. 2015

Archives of Biochemistry and Biophysics

View full text Add to dashboard Cite

Although a vaccine against hepatitis B virus (HBV) has been available since 1982, it is estimated that 600,000 people die every year due to HBV. An affordable oral vaccine could help alleviate the disease burden and to this end the hepatitis B surface antigen (HBsAg) was expressed in maize. Orally delivered maize material induced the strongest immune response in mice when lipid was extracted by CO2 supercritical fluid extraction (SFE), compared to full fat and hexane-extracted material. The present study provides a biochemical and biophysical basis for these immunological differences by comparing the active ingredient in the differently treated maize material. Purified maize-derived HBsAg underwent biophysical characterization by gel filtration, transmission electron microscopy (TEM), dynamic light scattering (DLS), UV-CD, and fluorescence. Gel filtration showed that HBsAg forms higher-order oligomers and TEM demonstrated virus-like particle (VLP) formation. The VLPs obtained from SFE were more regular in shape and size compared to hexane or full fat material. In addition, SFE-derived HBsAg showed the greatest extent of α-helical structure by far UV-CD spectrum. Fluorescence experiments also revealed differences in protein conformation. This work establishes SFE-treated maize material as a viable oral vaccine candidate and advances the development of the first oral subunit vaccine.

show abstract

“…Virions have properties that tend to aggregate (Gerba and Betancourt 2017), the aggregate status may be changed by adjustment of the pH or salt concentration and type of cationic salt in suspension (Mattle et al 2011). VLPs may also encounter the problem of aggregation due to their large size and complex structure, this can occur during the process such as during separation, purification and storage (Chen et al 2015;Jezek et al 2009;Kissmann et al 2008;Shi et al 2005).…”

Section: Discussionmentioning

confidence: 99%

Self-assembly into virus–like particles of the recombinant capsid protein of porcine circovirus type 3 and its application on antibodies detection

Wang

Duan

et al. 2020

AMB Expr

View full text Add to dashboard Cite

PCV3 capsid protein (Cap) is an important antigen for diagnosis and vaccine development. To achieve high-level expression of recombinant PCV3 Cap in Escherichia coli (E. coli), the gene of wild-type entire Cap (wt-eCap) was amplified from clinical samples, and three optimized entire Cap (opti-eCap) and one optimized Cap deleted nuclear location signal (NLS) (opti-dCap) gene fragments encoding the same amino acid sequence with wt-eCap were synthesized based on the codon bias of E. coli. Those gene fragments were inserted into the pET30a expression vector. One recombinant strain with the highest expressed soluble eCap from four entire Cap (one wt-eCap and three opti-eCap) and one recombinant strain expressed opti-dCap were selected for further purification. The purified eCap and dCap were identified by transmission electron microscopy (TEM), a large number of round hollow particles with a diameter of 10 nm virus-like particles (VLPs) were observed in eCap, whereas irregular aggregation of proteins observed in dCap. After formation the VLPs were applied as a coating antigen to establish an indirect ELISA (I-ELISA) for detection of PCV3-specific antibody in swine serum. 373 clinical swine serum samples from China collected in 2019 were tested utilizing the VLP-based I-ELISA method under optimized conditions. To the best of our knowledge, this is the first report of self-assembly into VLPs of PCV3 recombinant Cap. Our results demonstrated that the VLP-based I-ELISA will be a valuable tool for detecting the presence of PCV3 antibodies in serum samples and will facilitate screening of large numbers of swine serum for clinical purposes.

show abstract

Aggregation and antigenicity of virus like particle in salt solution—A case study with hepatitis B surface antigen

Cited by 30 publications

References 27 publications

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

Biochemical and biophysical characterization of maize-derived HBsAg for the development of an oral vaccine

Self-assembly into virus–like particles of the recombinant capsid protein of porcine circovirus type 3 and its application on antibodies detection

Contact Info

Product

Resources

About