ProMab Biotechnologies has years of experience providing CRO services with a skilled team of scientists who will work with you in developing your protein or peptide of interest. We offer protein expression services utilizing e. coli expression, baculovirus expression, mammalian expression, gene synthesis and peptide synthesis, as well.
Our high-level baculovirus expression service features:
Baculovirus expression takes 8 weeks from subcloning to expression in 3 phases starting at $5,000
- Competitive pricing
- High-level expression system
- Short timeline from cloning to protein recovery
Baculovirus expression systems have many known advantages including fast and cost-effective methods to generate large amounts of recombinant proteins in comparison to bacterial expression systems, particularly those requiring complex post-translational modifications. Our services include:
- Developing an effective strategy for molecular cloning and expression.
- Custom or high throughput cloning into a range of baculovirus transfer vectors.
- Small-scale transient expression screening to determine optimal expression conditions.
- Large-scale transient expression in shake flasks and/or WAVE bioreactor.
The insect cell/baculovirus expression vector system (BEVS) is becoming increasingly popular to produce recombinant proteins. The system offers several advantages,including:
- Able to perform complex post-translational modifications (PTMs)
- High success rate of soluble protein recovery
- Suitable to produce large protein complexes
- High protein expression levels compared to other higher eukaryotes
The ability of insect cell/BEVS to generate proteins with complex PTMs, coupled with high expression levels, makes it particularly suitable to produce mammalian proteins.
Transfection of sf9 insect cells
- Amplification/isolation of your gene of interest (GOI) and subcloning into a transfer vector with His-Tag or GST-tag.
- Sequence confirmation of insert.
- Transformation of baculoviral DNA transformed E. coli with the GOI to produce recombinant bacmid/GOI DNA.
Estimated time: 2-3 weeks
- Small-scale culture of E. coli and extraction of recombinant bacmid (recombinant viral DNA).
- Transfection of sf9 insect cells with recombinant bacmid to produce recombinant baculovirus.
- Small scale culture to test the expression level and subsequent optimization, if necessary.
Note: Net yield, purity, and solubility not guaranteed.
Estimated time: 3-4 weeks
Large Scale Transfection and Purification
- Virus stock preparation.
- Large scale culture and infection of sf9 insect cells for protein expression.
- Protein purification from 1 L of infected sf9 cells using affinity purification columns.
Estimated time: 3-4 weeks
- 5 μg of transfer vector
- recombinant virus
Yield ranges from 0.2– 50 mg/L of culture with average yield of 3 mg/L.