HyNPV Based Expression Services

HyNPV-Based-Expression-Services

As the principal BEVS viruses, AcMNPV and BmNPV each have their advantages as well as limitations. AcMNPV is easily cultured and infectious to a wide range of insects while BmNPV is suitable for industrial protein production, but the host is single.

To utilize the advantage of both AcMNPV and BmNPV viruses, Creative BioMart provides a "HyNPV expression system". Hybrid nuclear polyhedrosis virus (HyNPV) is the artificial hybrid virus of AcMNPV and BmNPV. In the HyNPV expression system, the protein expression is mediated by HyNPV rather than AcMNPV or BmNPV. The hybrid virus can infect the hosts of both AcMNPV and BmNPV.

Technology Description

HyNPV integrates the advantages of the two systems and avoids the disadvantages to some extent.

An AcMNPV helicase gene is added to the ORF 95 of BmNPV backbone to construct the HyNPV backbone. The changes of nucleotides and amino acids result in a host range expanded baculovirus expression system.

The working logic of the HyNPV follows the principle of Bac-to-Bac. In order to achieve efficient production of target protein, we suggest harvesting high titer recombinant HyNPV virus from the host insect cells of AcMNPV, and then these recombinant viruses can be used to infect silkworm or silkworm's cells.

HyNPV-Based-Expression-Services

Features of HyNPV Expression System

Compared to the parental expression systems that are extensively used, the HyNPV has the following advantages:

Expanded host range. The HyNPV is shown to be infectious to not only silkworm larvae, but also common cell lines (like Sf21, Sf9 and Tn-5).
Rapid recombinant protein production. That harvesting high titer virus from suspension cell culture system and then infect silkworm is more rapid than using any of the parental systems individually.
Suitable for industrial production. The hybrid system has a higher production efficiency than the parental expression systems by 50%—60%.
The HyNPV expression system will work more efficiently and enhance the stability of foreign expressed proteins making the hybrid system especially suitable for those proteins that are cysteine proteinase sensitive.

Service Content

Service Steps	Service Description	Timeline	Deliverables
System Selection （Optional）	Select donor vector Select host insect cell line	——	——
Gene Synthesis & Subclone Construction	Sequence design and codon optimization Gene synthesis Subclone the interest sequence to the donor plasmid	1-2weeks	Enzymatic / Sequencing validation report.
Recombinant Baculovirus Production	Preparation of recombinant Bacmid Transfect the insect cells with recombinant Bacmid Produce P1 Virus stock (low titer), P2 Virus stock (high titer), P1, P2 Western Blot and titer assay	2 weeks	Recombinant virus suspension WB report Titer test report
Small-Scale Expression & Purification	P2 virus transfect insect cell Protein purification SDS-PAGE, Western Blot	1.5 weeks	Target protein sample Quantitative test report
Optional Services	System optimization Secondary purification Tag removal Activity assay	1-2 weeks	Optimization solution Protein product Protein activity report
Large-Scale Expression & Purification (1-10L)	Culture amplification Western Blot Purification	1-3 weeks	Purified Protein QC report

Feel free to contact us or submit an online inquiry for help. Our customer service representatives are there to help you 7/24.

CONTACT OUR TEAM

Services