Insect Cell (nonlytic) Expression System

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:

Custom Service:



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


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%)