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 customers' requirements.
Protein Expression in Yeast cells is the most economical eukaryotic expression system for scale-up production of both secreted and intracellular proteins that require post-translational modifications, including humanized antibodies. Yeast-based production processes have been established with product yields in a multigram range. The organisms meet safety aspects in that they do not harbor pathogens, viral inclusions, or pyrogens. Alpha Lifetech Inc. provides customer a range of host yeast cells to meet the needs and different requirements of each project.
Host Yeast Strain:
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Saccharomyces cerevisiae |
Candida boidinii |
P. methanolica |
P. pastoris |
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H. polymorpha |
Kluyveromyces lactis |
Arxula adeninivorans |
Yarrowia lipolytica |
Alpha Lifetech Inc. deploys a wide range of expression vectors containing all E. coli elements for propagation in the E. coli system and an MCS for integration of yeast fragments like ARS, rDNA, selection marker, and expression cassette modules. All fragments are flanked by specific restriction sites (rDNA regions-BcuI/Eco47III, selection markers-Eco47III/SalI, expression cassettes-SalI/ApaI ARS-SacII/BcuI), in order to produce a high yield as possible.
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Fig. 1 model of the wide-range yeast vector system
Custom Service & Procedures:
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Phase |
Time |
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I. Gene Analysis and Synthesis |
2.5 weeks |
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-- Codon Optimization
-- Amplification/Isolation of target gene and subcloned target gene into an expression vector
-- Verification of the authenticity of the subcloned gene by restriction enzyme digest and sequencing.
-- Maxi-prep of recombinant vector DNA
-- Construct a linear expression vector |
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II. Transfection & Expression |
3 weeks |
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-- Transfection of recombinant constructs into a suitable yeast strain.
-- Verification of the authenticity of the subcloned gene by restriction enzyme digest and sequencing.
-- Expression optimization: medium, induction time, temperature and inducer concentration and so on.
-- SDS-PAGE and Western blot with tag-specific or custom-supplied antibody. |
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III. Small-Scale Culture |
4 days |
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-- 1L of yeast culture will be induced with inducer and harvested for protein purification. |
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IV. Purification and Purity Analysis |
1 week |
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-- Protein purification by one-step method, and purity analysis by SDS-PAGE or HPLC. |
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V. Endotoxin Control |
2 days |
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VI. Large Scale protein production & purification (optional) |
more than 4 weeks |
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-- Large-scale production up to 2,000 L (Employs fermenters and TFF large-scale filtration system) |
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Total |
7.5(or 11.5) weeks |
Service Features:
-- High success Rate: more than 95%
-- Fusion-Tag Selection: Single Tag or Combined Tags (His, Tag, GFP, FAM)
-- Traceability: Clear Genetic Information of Host Cells
-- Low Level of Endotoxin: <0.1 EU/100μg (LAL)
-- Low Level of Host DNA: < 1 ng/g
-- Fermentation Advantage: GMP condition, Various Scales (1, 5, 10, 30, 80L fermenter)
-- Time & Products: Short Time Cycle; High purity (no less than 95%)
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