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Quick List | |
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Price and timeline Service Terms Confidentiality Delivery Request to the Customers Why SinoBio Why Outsourcing How to order Frequently Asked Questions | |
Introduction
SinoBio Biotech provides diverse host strains for your
protein expression such as DH5a, BL21, BL21 lys, Rose, Star, JM109. The
different feature of such strains may help you find out better host. Different
promoter in addition to different expression vectors for your choice. We highly
recommended try expression vector with different promoters and fusion tag to
boost your target protein expression. The promoters can be T7, Tac, Cold, T3,
PRPL. Diverse fusion tags are available to facilitate your protein expression
and purification. The tags include TRX, HIS, GST, NUS, SUMO, FLAG, MYC. These
tags can be included in your target protein or be removed with a protease after
purification to get a gutless protein.
Price and timeline
| Sequence design | 1 work days | free |
| DNA synthesis | 2 week | 1$/bp |
| Clone to expression vector | 1 week | Free or 300$/cst |
| Expression strain construction | 2 work days | |
| Expression Screen | 2 work days | |
| Large scale expression | 3 work days | |
| Large scale purification | 2 week | |
| Conventional refolding to ensure solubility | 3 work days | |
| Protease digestion to remove the tag | 2 work days | |
| After digestion refolding when necessary | 2 work days | |
| Fine purification to get final product | 1 week | |
| Quality analysis for final product | 3 work days | |
| Interim report and final report | 1 work days | |
| Shipment | 3-5 days |
Service Terms
A
general full service includes the following steps:
1. Choose of proper expression systems, by the customer
or discussing with the customer.
2. Choose of proper
expression vectors, by the customer or discussing with the customer.
3. Sequence optimization for the exact expression systems, by the customer or
discussing with the customer, or a native resource is used under customers'
requirement.
4. Confirm the order by the customer via
email or fax.
5. The target gene will be cloned into the
proper expression vector you desired. Subclone into one of the expression vector
will be free if the synthesis is provided by SinoBio.
6.
Sequence validation.
7. Expression screen.
| Expression optimization will include: | ||
 |
culture medium |
 |
| induction condition | ||
| solubility of target protein | ||
| Purification procedure will include: | |
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Affinity purification |
| IEX; HIC; SEC purification | |
| Remove of tag part when required | |
| Conventional refolding to ensure the protein solubility when necessary | |
| Refolding screen when required | |
| Endotoxin control when required | |
9. Quality control and confirm with the customer for the
final products.
10. Shipment.
*
Interim reports will send to customers for the synthesis, expression screen and
purification, or a rotun reports for every two week, and all expectance will be
reported to and discussed with the customer immediately.
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 e.coli 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 from
expression screen is 15 business days. 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'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.
Correlative Service
Gene Synthesis
Polyclonal Antibody Production
Protein Analysis(HPLC, LAL, AA Sequencing, MS, MS/MS)
Protein PEGylation
Protein Refolding
Activity assay
for some of the proteins
Expression in
P.Pichia
Expression in
Insect Cells
Expression in
Mammalian Cells
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.
Why You Need Outsourcing
Not all the protein needed are commercial available.
Nowadays, many proteins are physically available from commercial suppliers.
However, there are a significant number of proteins that exist only as digital
sequences. Also, Biological and Pharmaceutical research require proteins of
man-made and predicted sequences that do not exist naturally. Custom protein
manufacture provides a unique approach to obtain these otherwise unavailable
proteins.
CMO can provide you special protein sequence under your
requirements, Point mutation, insertion and deletion, fusion to different tag,
In addition to non-commercial available proteins,
custom protein manufacture can also provide you at least the following features:
.Addition of elimination of protein modification sites such as phosphorylation,
glycosylation or Pegylation sites
.Division of genes into modules or cassettes for protein domain swapping,
insertion, deletion or duplication
.Introduction of specific mutations within a particular gene module
.Chimeric/fusion protein production
CMO can provide different quality protein meet your
requirements.
Lower purity to reduce your cost esp. when using as
antigen.
Higher purity to meet
your special application, e.g. crystal analysis
Even a large amount of proteins are commercial available, your request may have
some special feature such as different purity requirements. If you need small
amounts of these proteins, commercial suppliers may be more suitable with more
insurance.
CMO can reduce your cost of money as well as time
Protein expression and purification is still an art of skill. Not a theory
can give a promised suggestion of how to get your desired protein. SinoBio
Biotech have more than 300 recombinant protein manufacture experience, developed
about 40 full qualified cytokines and tens of biological enzymes. Experience
isn't a promise but can really improve the possibility of success.
Weeks of time may be spent for cloning a full-length cDNA, and another couple of
weeks construction and expression test, purification and quality analysis. Just
thinking about the salary you have to pay for a simple protein, no matter the
cost for reagents you spent.
SinoBio Biotech, using our
skilled techniques, the whole process can shorten to ONE month. If the
expression strain is provide, 2-3weeks can get the aim where you want go.
CMO can reduce your adventure of fail
For protein purification, we promised no charge if we failed to get the
protein, no matter the reason is no expression or difficult to be purified. By
this means, we take the adventure of a failed project, at least the time your
scientist may wasted on it if it's impossible to be expressed or purified.
CMO/CRO as the first choice for your target protein:
Large amount
Long term usage
No
commercial available
Special requirements such different
purity
CMO/CRO as the secondary choice:
Limit amount required, such as microgram to milligram
Low cost product available from commercial providers
Biological activity must be insured.
How to
order
Please download the order form to specify your information, billing
information, and project information;
Please email the
completed order form to info@sinobio.net
Please follow these instructions closely to speed up
your ordering process.
1. SinoBio Biotech adopt a non-disclosure policy to all
of our customers. To protect your proprietary information please mail, email
(info@sinobio.net) or fax (+8621-5079-8029) your confidentiality agreement to us
for sign. If intended, this should be done before any information is released to
us. If you don't have your own NDA/confidentiality agreement, you can find ours
at here. Feel free to modify it to suit your specific needs.
2. Call us (+8621-5079-8029) or email us info@sinobio.net, your gene/protein
sequence and particular manipulations you wish us to perform. Particularly give
clear instructions on the following options: codon optimization vs. no
optimization; host organisms; single host vs. multiple host optimization. Total
amount of final protein you required, the qualities and QC terms you required.
Frequently Asked Questions: E.coli Expression Systems
1) What's the possibility of my protein expressed in e.coli?
2) Should I adapt the codon usage to e.coli favioured?
3) What's the size limit of foreign protein expression in e.coli?
4) Should I introduce a fusion tag to my protein?
5) What should I do next when I found my construction not work?
6) Should I try secret expression in e.coli?
7) What quality of the protein SinoBio Biotech will provide? What should I required?
8) What should I do if the protein expressed in inclusion body?
1) What's the possibility of my
protein expressed in e.coli?
To express a foreign gene in e.coli still a kind
of challenges. According to our experiences, only one third of native
sequence of mammalian genes can be expressed directly in e.coli. By
fusion to different tag, about half of non-expressed protein can be
expressed.
2) Should I adapt the codon usage to e.coli
favioured?
Codon usage in e.coli is quite different from
eukaryocytes. To adapt the condons to e.coli preferred can improve the
expression level in most case. The CAI program should be one of the recommend
tools for optimize your gene sequence to facilitate expression in e.coli.
3) What's the size limit of foreign protein
expression in e.coli?
A gene at the range of 200bp-3000bp can be tried in e.coli. Too large or
small proteins are not suitable to be expressed in e.coli unless there are
references to do so.
4) Should I introduce a fusion tag to my
protein?
Fusion expression may be the best way to improve the possibility of protein
expression. By fusion with Trx Tag, we found at least half of non expression
genes can be expressed well in e.coli. The fusion tag can be removed by protease
digestion. Enterokinase should be the first choice for the better specificity.
A fusion tag can also improve the solubility of your
target protein to reserve the biological activity.
5) What should I do next when I found my
construction not work?
Change to different promoters but not the different vectors with same
promoter are more safe way to get your protein expressed. Sinobio Biotech will
provide diverse choices of expression promoters in e.coli such as T7, Tac, PRPL
and T3 promoter.
6) Should I try secret expression in e.coli?
Secret expression is not recommended because we found in most case the
e.coli cannot process the signal peptide correctly. Of course there are still
lots of cases for secret expression in e.coli esp. for small proteins.
7) What quality of the protein SinoBio
Biotech will provide? What should I required?
Keep in
mind that the qualities of your protein should be sufficient is enough. For
crystal analysis, 90% should be ok but some times the purity should be higher
than 95%. But if you just want the protein as a antigen, 70-80% is enough and
contains a tag can be allowed.
If you just want to get
the protein for immunolize animals to produce antibodies, inclusion bodies
should be one of the good choice as well as contains a small purify tag such as
HIS6.
The same goal of service provider and the customer
is to get the suitable protein for customer's usage. The simple the better for
it reduces your cost and improve the possibility of success. We are welcome to
any challenge from invests and strategy cooperators but not the customers.
8) What should I do if the protein expressed
in inclusion body?
Inclusion bodies can be renatured by different in vitro refolding process.
There a lots of documents tell you how to refolding a protein, but none of them
is promised. Generally, a 10% refolding efficiency is acceptable for most
proteins. SinoBio Biotech provides a systematic refolding strategy for you to
decide the optimized refolding parameters, such as the refolding pH, redox, and
different addictives.
