Virus like particles (VLP) Expression platforms

Structure of capsid proteins is important to stimulate the immune response and it is depended on the post translational modification of different expression systems. The various expression systems such as prokaryotic (bacteria, yeast) or eukaryotic (baculovirus/insect cell, mammalian cell and plant) have been used to produce the VLP.  In addition, cell-free expression systems have also been used successfully. Selecting a suitable system is important and system has both advantages and disadvantages (Fuenmayor et al. 2017).


1. Bacteria based virus like particles (VLP)

Bacteria are one of the most widely used systems for the non-enveloped virus like particles production. Escherichia coli-based expression system has advantages such as cost-effective, rapid cell growth and high protein expression level to produce safe VLP. However, prokaryotic expression system is not suitable for some protein due to the lack of PTM system. Various VLP vaccines produced by E. coli have been successfully used against infectious and non-infectious diseases.



2. Yeast based virus like particles (VLP)

Yeast is also most commonly used for VLPs production. Saccharomyces cerevisiae and Pichia pastoris, are the widely used yeast expression system. The yeast system has the advantages such as rapid cell growth, high yield, scalability, cost-effective, and PTM process.



3. Baculovirus/Insect cells (B/IC) based virus like particles (VLP)

The B/IC expression system is used to produce both enveloped- and non-enveloped-VLPs. The insect cell lines viz. Spodoptera frugiperda (Sf9/Sf21) and Trichoplusia ni (Tn5) is commonly used for VLP production. Due to its fast growth, it is convenient to produce the vaccines for outbreaks such as influenza virus. It has advantages such as high yield of proteins, complex PTM pathways. The Baculovirus/insect cell platform has been used to develop the vaccine against HIV 1, influenza virus A, Chikungunya virus, severe acute respiratory syndrome (SARS), Ebola virus, dengue fever virus, Rift Valley fever virus (RVFV), Norwalk virus and HCV.



4. Plants cells-based virus like particles (VLP)

Plant-based VLP platform is a versatile and cost effective. It has the advantages such as expression levels of up to 80% total soluble protein, low refining costs, and high-performance expression processing. Numerous studies on the VLP vaccines based on plant system have been effectively created. The plant viruses such as Tobaco Mosaic Virus (TMV), cucumber mosaic virus (CMV), alfalfa mosaic virus (AIMV), cowpea mosaic virus (CPMV), papaya mosaic virus (PapMV), and the potato X virus (PVX) have been used for the VLP platform.



5. Mammalian and avian cells-based virus like particles (VLP)

Animal cell expression systems is a valued platforms for producing the structural proteins of non-enveloped and enveloped VLPs. Animal cell platforms are most effective due to its complex and precise PTMs. The animal cell lines such as CHO, baby hamster kidney-21 (BHK-21), human embryonic kidney 293 (HEK293), CAP‐T cell line derived from human amniocytes, Vero 9, and east lansing line-0 (ELL-0) are widely used system for the VLPs production. Among them, CHO has been most commonly used and it has the advantage over other cell lines.



Table 1. Advantages and limitation of different expression system
Expression system Advantages Disadvantages
Bacteria Cost-effective
Scalable
Fast growth
High level of expression
Genetically stable
Poor immunogenic
Absence of PTM
Poor protein stability
Presence of bacterial endotoxin
Yeast Low cost
no endotoxin contamination
high density fermentation
support some protein folding
Low yield than bacteria
Lack of mammalian PTM system
High mannose modification
Insect cells Uptake and deliver large amount of DNA
High level of protein expression
Support PTM
Proper protein folding and assembly
Difficult to scale up
Production cost is high
Baculovirus contamination possibility
PTM not similar to mammalian system
Incomplete protein modification
Mammalian cells Proper PTM, protein folding and assembly Large scale production
Expression time
Low yield
Mycoplasma contamination
Plant cells Highly scalable
Cost effective
High expression
Proper folding and assembly
Lower yield than mammalian cells
Technical issues


Picture loading failed.View the Knowledge base of virus like particles (VLP):

    - Virus-like particles (VLP) Platforms for Developing immunogens, vaccines and drug carriers
    - What is virus like particles (VLP)?
    - GM VLPx™ Virus-like particles development platform
    - Virus like particles (VLP) Expression platforms
    - Virus like particles (VLP) as immunogens/antigens of transmembrane protein (TM)
    - Virus like particles (VLP) for drug delivery
    - Virus like particles (VLP) as vaccine
    - Virus like particles (VLP) for drug discovery





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