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Escherichia coli (E. coli) provides for a relatively simple, fast, inexpensive and robust system for native and heterologous recombinant protein production. While unable to render many of the complex eukaryotic post-translational modifications, technologies such as co-expression of chaperones/kinases/rare tRNAs and engineering of strains that provide a pro-disulfide bond (oxidative) environment hallmark the technological progress achieved in the E. coli system over the last two decades. abm recommends the E. coli system when post-translational modifications such as glycosylation are not crucial to the intended downstream application(s).

The mammalian expression system provides the only authentic and feasible option in cases where the protein of interest needs to be embellished with characteristic, mammalian post-translational modification(s) and/or entails a complex translational machinery for proper folding and targeting (such as membrane proteins). Therefore, abm recommends its mammalian expression service for proteins for which the native folding and post-translational modifications are of a complex nature (beyond the purview of E. coli) and indispensable to the customer’s intended application(s).

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Bacterial Expression
Mammalian Expression

“I am a fan of abm: great product quality, great price, great rewards and great experience.”

Dr. Vincent Emond, Centre de recherche du CHU de Québec CHUL

Service Details

 

Custom protein production in E. coli

PHASE I SERVICES CAT. NO. UNIT
  • Codon optimization (where applicable) and gene synthesis (up to 4kb^) of sequence provided by customer.

^Minimum charge of $99.00, complex sequences are subject to additional charges

C098 1 bp
  • Cloning/sub-cloning# of gene of interest into a protein-encoding gene expression vector
  • Sequence confirmation

#If you intend to submit your own insert, please provide vector map and sequence information for compatibility review. Inserts with restriction sites that are not compatible are subject to an additional $100 charge.

C101 1 Service
  • A set of multivariate small scale protein production trials are conducted to test for yield, solubility and optimal growth conditions
C102 1 Service
HASE II SERVICES
  • Large scale recombinant protein production and purification (purity ≥85%)
C103 1 Service
ADD-ON SERVICES
Proteolytic removal of affinity tag C104 1 Service
Special Buffer Request C118 1 Service
Special Concentration Request C119 1 Service
Lyophilization Request C125 1 Service
Endotoxin testing and removal (<1EU/µg of protein) C194 1 Service

 

Deliverables:

  1. If only Phase I is completed (i.e. up to C102), a service report will be provided as the only deliverable. All charges in Phase I will still be applicable.
  2. If both Phases I and II are completed, deliverables will include a service report and the protein only.
  3. If technical difficulties prevent successful tag removal (Cat# C104), all quotation charges will still apply except the Cat# C104 fee.

Note:

  • Phase I: Full payment required to initiate Phase I
  • Phase II: A 30% non-refundable deposit is required to initiate phase II

Custom protein production in Mammalian cells

PHASE I SERVICES* CAT. NO. UNIT
  • Codon optimization (where applicable) and gene synthesis (up to 4kb^) of sequence provided by customer.

^Minimum charge of $99.00, complex sequences are subject to additional charges

C098 1 Service
  • Cloning/sub-cloning# of gene of interest into adenoviral expression system
  • Sequence confirmation
  • Viral Packaging

#If you intend to submit your own insert, please provide vector map and sequence information for compatibility review. Inserts with restriction sites that are not compatible are subject to an additional $100 charge.

C110 1 Service
  • Small scale expression trial verified by Western Blot
C111 1 Service
PHASE II SERVICES
  • Large scale recombinant protein production if adequate levels of expression is deemed in Phase I
C112 1 Service
  • Large scale recombinant protein purification (≥85% purity)
C113 1 Service
ADD-ON SERVICES
Proteolytic removal of affinity tag C114 1 Service
Special Buffer Request C118 1 Service
Special Concentration Request C119 1 Service
Lyophilization Request C125 1 Service

 

Deliverables:

  1. If only Phase I is completed (up to C111), a service report will be provided as the only deliverable. All charges in Phase I will still be applicable.
  2. If both Phases I and II are completed (up to C113), deliverables will include a service report and the protein only.
  3. If technical difficulties prevent successful tag removal (Cat# C114), all quotation charges will still apply except the Cat# C114 fee.

Note:

  • Phase I: Full payment required to initiate Phase I
  • Phase II: A 30% non-refundable deposit is required to initiate phase II

Additional Info

  1. Disclaimer for intended use: All of abm's recombinant proteins are for research use only and not intended for therapeutic/diagnostic applications. abm is not liable for any violation (and associated repercussions) to the specified use of its entire suite of recombinant proteins.
  2. Disclaimer for 'E. coli' expression system: The choice of utilizing a prokaryotic expression system (Escherichia coli) for eukaryotic protein production is solely up to the discretion of the customer with respect to being aware of the limitations to the rendition of correct number and type of Post Translational Modifications (PTMs).
  3. Disclaimer for extra amino acids: abm does not guarantee that the recombinant protein product will be free of any additional amino acids residues. Cloning may lead to the insertion of some amino acids at either and/or the N and C-terminus of the recombinant protein product which in most cases, is innocuous to the stability/functionality of the recombinant protein.
  4. Disclaimer for turnaround time: Note that the turnaround time indicated above is our best estimate currently. Final turnaround time is subject to change depending on actual experimental conditions.
  5. Disclaimer for functionality: abm is not in a position to guarantee protein functionality.

  1. Disclaimer for intended use: All of abm's recombinant proteins are for research use only and not intended for therapeutic/diagnostic applications. abm is not liable for any violation (and associated repercussions) to the specified use of its entire suite of recombinant proteins.
  2. Disclaimer for extra amino acids: abm does not guarantee that the recombinant protein product will be free of any additional amino acids residues. Cloning may lead to the insertion of some amino acids at either and/or the N and C-terminus of the recombinant protein product which, in most cases, is innocuous to the stability/functionality of the recombinant protein.
  3. Disclaimer for turnaround time: Note that the turnaround time indicated above is our best estimate currently. Final turnaround time subject to change depending on actual experimental conditions.
  4. Disclaimer for functionality: Note that abm is not in a position to guarantee downstream functionalities.
  5. By default, a His tag will be used for purification purposes. Otherwise, additional charges apply and please provide a verified gene expression antibody along with a positive control.
  6. Charges for C112 and C113 determined after customer has reviewed report from C111 and requested for Phase II. An estimate can be provided during quotation process.

Documents

Custom Protein Production Quotation Request Form

FAQs

You may find a list of our protein expression system backbones at the below link: https://www.abmgood.com/Custom-Cloning-Service.html

Custom protein production in Mammalian cells: Please note that only the recombinant protein is delivered when Phase I ---> IV are ordered. If you would like to receive the product resulting from each phase, you may do so at an extra charge: Phase II: Stable protein producing cell-line- polyclonal (293 cells) @ $2000 Phase III: 500ul of crude lysate (in WB sample buffer, denatured) @ $100 You may choose to stop at any phase, however please note that this is subject to material and labor costs incurred thus far. Handling charges apply if you wish to receive the deliverables from a specific Phase.

The protein will be purified without any denaturing steps but we cannot guarantee whether the protein is folded properly within the E. coli system.

We require customers to submit inserts that can be subcloned into our protein vectors to initiate phase I. Please provide your vector map and sequence for us to evaluate compatibility. If you are unable to provide the insert, we can provide an additional gene synthesis service. Please provide the sequence for this additional service.

Citations

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