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Terms and timeline

Phase 1. Cloning of the target gene into a baculoviral expression vector
    Amplification and isolation of the gene and subcloning it into a pFastBac donor plasmid.
    Verification of the authenticity of the subcloned gene by restriction enzyme digest and sequencing.
    Maxi-prep of the recombinant vector DNA.
Time: 2~3 weeks
Provide to customers: Correct pFastBac donor plasmids and sequencing results.

Phase 2. Construction of recombinant Bacmid
    Transformation of recombinant pFastBac donor plasmid into DH10Bac E. coli cell.
    Antibiotic selection of E. coli containing recombinant Bacmid.
    Mini-prep of high molecular weight DNA (recombinant Bacmid DNA).
Time: 1 week
Provide to customers: Recombinant Bacmid DNAs and test results.

Phase 3. Transfection of insect cell and amplification of baculoviral stock
    Transfection of insect cells (Sf-9).
    Collection of recombinant baculovirus particles and determine viral titer.
    Plaque-purification of baculovirus if desired.
    Increase viral titer and amplify virus.
    Test for expression of the recombinant protein by western blot.
Time: 3 weeks
Provide to customers: Recombinant baculoviral stocks and result reports.
Note:
1. If we do not get the baculovirus (failed in transfection), we would not charge for this phase. If we successfully get the baculovirus but the test result shows no target protein expression, we will only charge 700 USD each in this phase.
2. Customer must provide the antibody for Western-blot if the test result is required.

Phase 4. Recombinant protein expression and purification
    Infection of 1 liter of insect cell (Sf-9, Mimic Sf-9 or High Five) for recombinant gene expression.
    Harvest of expressed target protein
    Protein purification by a variety of columns including affinity, gel filtration, ion exchange and hydrophobic column.
    Provide at least 0.5mg target protein at soluble conditions. The purity of the protein is above 80%.
    Test for the result of each step by SDS-PAGE or Western blot (if desired).
Time: 4~6 weeks
Provide to customers: Qualified target protein and purification result report.
Note: Customer must provide the antibody for Western-blot if the test result is required.


Confidentiality
    Your confidential sequence to us by mail will be covered. We will sign non-disclosure or confidentiality agreements when necessary.
    A general template for MTA and CFA are available and you may change the terms according to your requirements and discuss it with SinoBio.

Delivery
    Materials delivered to customers include the following:
    1. Baculovirus stock.
    2. DNA sequence data (When necessary.).
    3. Required final products in conventional buffer.
    4. Manufacture reports, including the major process, interim results, quality control.
    Delivery time:
    The typical delivery time is 6 weeks. Time for shipment is about 4 business days. This may vary depending on where the customer located and accidentally, delayed by custom check.
    The final products will be shipped via fedex. DHL is optional according to the customer£¦#8217;s requirements.

Request to the Customers

    1. Tell us the target DNA sequences or Genbank accession number or protein sequence through the E-mail.
    2. Provide us the requirements of the final products essentially the purity and amount, other quality control requirements are optional with additional charge such as HPLC, AA Sequencing, MS, MS/MS, biological activity, endotoxin control etc. For details please read protein analysis.
    4. The customers need to pay for 50% fees as subscription for the first order.
    5. For more genes to be expressed or complex project, please contact us for a quote.

Why SinoBio
    Professional: SinoBio Biotech focus on recombinant protein manufacture only, and the focus makes us more professional. All of the group leaders are come from big biopharmaceuticals with more than 10 years experience on protein production. The company are well equipped with Bioengenering fermentor, AKATA, HPLC and we can provide cGMP standard for your production. The fermentation scale is from liters to 500 liters. Final production is scale from milligram to tens of gram.
    Expertise: SinoBio's protein experts have more than 300 different protein purification experience, involved in NINE biologics development, most of them are in market and some of them in clinical trails. They also developed about 40 qualified cytokines and tens of biological enzymes.
    Quality: Diverse quality control method available for your choice such as amino acid sequencing, MS, MS/MS, LAL test, HPLC in addition to conventional SDS-PAGE analysis.
    Speed: Fast turnaround time (~10 days for a basic order)
    Price: Competitive prices as low as 800$
    Confidentiality: Strictly confidential customer's data. SinoBio will sign non-disclosure or confidentiality agreements when needed.