Protein production and purification in a human model
CliniSciences offers specialized services in recombinant protein production and purification using human HEK293 cells, ensuring the incorporation of authentic post-translational modifications (PTMs) critical for protein functionality. HEK293 cells are renowned for their ability to perform complex PTMs, such as glycosylation, phosphorylation, and proteolytic cleavage, which are often absent in prokaryotic systems like E. coli, making them ideal for producing biologically active proteins.
Principes
The human HEK293 cell line provides a robust mammalian expression system capable of performing a wide array of PTMs essential for proper protein folding, stability, and activity. These modifications, including glycosylation, phosphorylation, and proteolytic cleavage, are critical for the biological function of proteins in both in vitro and in vivo applications. Unlike prokaryotic systems, which lack the cellular machinery for complex PTMs, HEK293 cells closely mimic human physiology, producing proteins with human-specific modifications that enhance their suitability for therapeutic, diagnostic, and research purposes.
Recent advancements in transient transfection methods, such as optimized PEI-based protocols, have further improved expression efficiency in HEK293 cells, enabling high yields and scalability for biopharmaceutical applications.
Service Components
Our comprehensive service for recombinant protein production in HEK293 cells includes:
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Gene Synthesis and Codon Optimization: Designing and synthesizing the gene of interest (GOI) with codon optimization to enhance translation efficiency and protein yield in HEK293 cells.
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Gene Cloning and Vector Construction: Inserting the GOI into a high-efficiency expression vector, incorporating tags (e.g., His-tag, Strep-tag) for streamlined purification and detection.
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Transient Transfection and Expression: Transfecting HEK293 cells using optimized transient or stable methods to maximize protein synthesis, with scalability for high-throughput production.
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Protein Purification: Harvesting culture supernatant or cell lysate, followed by affinity chromatography (e.g., Ni-NTA, Strep-Tactin) and optional size-exclusion or ion-exchange chromatography for high purity.
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Quality Control and Characterization: Analyzing protein purity and integrity via SDS-PAGE and Western blot, with optional mass spectrometry for molecular weight and PTM verification.
Advantages
CliniSciences stands out in recombinant protein production through:
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Expertise in HEK293 Systems: Deep specialization in human cell lines ensures proteins with authentic human-specific PTMs, ideal for biopharmaceutical applications.
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End-to-End Solutions: Comprehensive services from gene synthesis to protein characterization, streamlining the production process.
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Customization and Flexibility: Tailored solutions, including tag removal, endotoxin reduction, and scalable production to meet specific project needs.
Request a Proposal from our Technical Support Team to tailor our protein production services to your needs.
Related Services
In addition to HEK293-based protein production, CliniSciences offers:
Related services available :
=> Gene synthesis
=> Gene cloning
=> Polyclonals production
=> Monoclonals production