Discovery Biology Services
Protein Expression and Purification

There are four gene engineering expression systems in Viva, including escherichia coli expression system, baculovirus expression system, protein expression with suspension mammalian cells and yeast expression system. Through a complete protein expression system, we can provide customers with efficient protein expression and purification services for the study of protein structure and function.

Service Content
  1. Plasmid Construction

    Viva Biotech provides customized gene cloning services, including codon optimization, gene synthesis (outsourced) and subsequent cloning. After you provide a gene template, send a target vector and cloning demand, or provide a protein sequence and nucleic acid sequence, we will provide existing or customized services according to the specific specifications required. In addition, we can provide one-stop service for high yield protein expression, purification and production.

     

    · Subclone service

    · Site directed mutagenesis cloning service

    · Missing, insert mutant cloning service

    · Plasmid extraction (Plasmid Endofree Miniprep, Midiprep, Maxiprep, Megaprep, Gigaprep)

    · Plasmid endotoxin removal and determination service

  2. High Yield Protein Expression

    Escherichia coli expression

    · The constructed plasmid can be transformed into a variety of commercial strains (such as BL21(DE3), BL21 (DE3) Gold, BL21 (DE3) CodonPlus RIPL, and Rosetta)

    · Inducible expression at different temperatures (16℃- 37℃) can be performed

    · Different culture mediums (LB, TB, 2xYT, M9) can be selected

    · Different induction methods (IPTG induction, arabinose induction, autoinduction) can be selected

    · Coexpression chaperone can be selected to improve soluble expression

    · High throughput expression test based on 96 well plate (which can test the expression differencesof different strains and different constructs)

    · Expression of isotope labeled proteins (including 15N labeling, 15N13C double labeling, 15N2Hdouble labeling, 15N13C2H triple labeling, and 13C-Me ILV selective labeling based on 15N2Hdouble labeling)

     

    Baculovirus expression system

    · Generationand amplification of Baculovirus

    · Baculovirus systems: Bac-to-Bac system; Bacmam system; Linearized co-transfection systems such as pVL1392, ProEASY, BD BaculoGold system, pTriEX …

    · Optimization of expression: cell lines, infection time, infection ratio (MOI), expression temperature…

    · Capacity of large-scale expression:>1000 L/month

    · Capacity of construct numbers: >200/month

    · Preparation, preservation and optimization expression of BIIC (Baculovirus Infected Insect Cell)

    · Insect cell transient experiment (InsectSelect BSD System)

     

    Protein expression with suspension mammalian cells

    · Transient transfection expression and BacMam infection expression services in a varietyof suspension cell lines including Expi293F, ExpiCHOS, Freestyle 293F, and Expi293F™ GnTI-cell

    · Cell culture service with suspension mammalian cell including transient transfection, viral infection, and large-scale expression with stablecell line culture up to 400 L/month

    · Stable cell line generation for protein expression

     

    Yeast expression system

  3. Protein Purification

    · Inclusion body purification

    Inclusion body/Insoluble protein is expressed, washed followed by denatured and dissolved, and then purified and refolded (or purified after refolding) to obtain soluble protein

     

    · Secretory protein purification

    The signal peptide is used to guide the expression of secretory protein outside the cell, and the cell supernatant (culture medium) is collected directly for purification

     

    · Common modifications involved in purification

    Biotinylation (in vivo or in vitro), phosphorylation, dephosphorylation, deglycosylation

     

    · Enzyme digestions involved in purification

    TEV, Ulp1, PreScission (or 3C), Thrombin, and Ek

     

    · Other technologies involved in purification

    Crosslinking, Nanodisc, and Fibrillization

  4. Protein Production

    · Production of soluble protein

    · Production of renaturation protein

    · Production of secretory protein

    · Membrane protein assembly

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