Alpha Lifetech Inc. focuses on recombinant protein expression in different Biosystems for many years, including prokaryotic and eukaryotic organisms, as well as the development of novel high-potency recombinant proteins to meet your technical, program, and budget requirements.
Protein expression based on baculovirus-infected cells has become the preferred choice for many applications because posttranslational modifications and correct disulfide-bond formation are necessary for protein folding and activity. The proteins produced in insect cells involve mainly two types of host cells: Sf9, Sf21, High5, S2 and S3 cells.
The basic procedures of protein expression in Nonlytic Insect Cell System:
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Custom Service:
Phase |
Time |
I. Plasmid Construction |
3 weeks |
-- Gene Analysis/Amplification
-- Subclone Design: non-excrete/excrete plasmid, tag selection
-- Integration: Target gene into the vector
-- Verification of the authenticity of the subcloned gene by restriction enzyme digest and sequencing
-- Maxi-prep of recombinant vector DNA |
|
II. Transient Gene Expression |
1 week |
-- Transfection of host cells with the recombinant plasmid.
-- Culture condition optimized: Induce time, Medium. |
|
III. Stable Expression Cell Line (option) |
3-6 weeks |
-- Transfect cells with plasmid and select living cells from a screening condition -- Single-cell cloning: Dilute cells into single ones and culture them in a 96-well plate
-- Fermentation or cell factories culture |
|
IV. QC Verification & Small-scale Amplification |
1 week
|
-- Validation for expression of the recombinant protein by SDS-PAGE (Customer must provide appropriate antibodies, if desired WB)
-- Generation of 500 ml of high-density cell harvest |
|
V. Purification |
1 week |
-- Protein purification using one-step method: affinity chromatography (Ni-NTA/glutathione column) -- Validation for expression of the recombinant protein by SDS-PAGE |
|
Total |
9-13 weeks |
Service Features:
-- High Success Rate: more than 95%
-- Fusion-Tag Selection: Single Tag or Combined Tags
-- Traceability: Clear Genetic Information of Host Cells
-- Fast Expression of recombinant proteins
-- Low Level of Endotoxin: <0.1 EU/g (LAL)
-- Low Level of Host DNA: < 1 ng/g
-- Fermentation Advantage: Various Scales (1, 5, 10, 30, 80L fermenter)
-- Time & Products: Short Time Cycle; High purity (no less than 95%)