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Phase 1. Cloning of the target gene into a CHO or HEK 293 cell expression vector.
    Amplification and isolation of the gene and subcloning it into a CHO or HEK 293 cell expression vector.
    Verification of the authenticity of the subcloned gene by restriction enzyme digest and sequencing.
    Midi-prep of the recombinant vector DNA.
Time: 2~3 weeks
Provide to customers: Correct expression vector and sequencing results.

Phase 2. Small-scale transfection and detecting the expression with Western-blot. (Customer should provide the antibody)
    Transfection of CHO or HEK 293 cell with Lipo2000 reagent.
    Detection of the expression in every 12 hours from 24 hours to 72 hours after transfection.
Time: 1 week
Provide to customers: Expression detection result report.

Phase 3. Selection of the high expression stable cell line. (Optional)
    Transfer the cell aggregate to 96 well plate, for about 2 to 3 groups per well, and then add cell culture medium which contained MTX.
    When the clone cell groups grow up to about 2/3 well, sample the supernatant and detect the expression.
    Pick up the high expression cell group for next selection, and also increase the concentration of MTX in the cell culture medium.
    Repeat the step 2 and 3, using this press selection method to increase the expression amount until finally establish the cell line.
Time: 6~12 week
Provide to customers: High expression stable cell line and result report.

Phase 4. Pilot-scale protein expression and purification.
    1 liter of CHO or HEK 293 cell was cultured for protein expression.
    Transfection of cells with Lipo2000 reagent and expression of the target protein (if not establish the stable cell line).
    Collection and concentration of the expressed protein.
    Protein purification by a variety of columns including affinity, gel filtration, ion exchange and hydrophobic column.
    Polish of the purified protein include endotoxin removing and sterile filtrating.
Time: 6 weeks
Provide to customers: Qualified target protein and purification result report.

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. Expression strain in glycerol.
    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.