Fc Fusion Protein Production Services

Syd Labs has extensive experience with making DNA constructs for Fc fusion protein production, expressing and purifying a number of Fc fusion proteins from microgram to kilogram scales in mammalian cells, such as CHO and HEK 293 cells. With improved DNA construction system and fully optimized secretion signal and linker sequences, Syd Labs can efficiently complete a project from DNA construction and transient production of Fc fusion proteins in CHO and HEK 293 cells, to Fc fusion protein purification. N-term or C-term of Fc fusions of peptides and proteins in secreted form as well as multiple Fc backbone are available upon request.

The Fc region (the antibody fragment crystallizable region) brings a variety of benefit to the fused proteins and peptides. Fc fusion allows simple and efficient purification with protein A and/or protein G, and accurate quantification and capturing in receptor-ligand affinity measurement. Fc fusion significantly improves pharmacokinetic property of the fused proteins and peptides because Fc can escape proteolytic degradtion. Fc can increase the plasma half-life of Fc fusion proteins due to interaction with the neonatal Fc receptor (FcRn) and the slowing of renal clearance due to their larger size. Because they are homodimers, Fc fusion proteins may have improved apparent affinity to binding partners due to increased avidity. Because many receptors are only functional in a dimeric form, the dimeric receptor-Fc fusion protein can mimic the activated form and enhance its affinity for the cognate ligand. With human-specific glycosylation, Fc chimeric proteins can be used in studies involving human proteins and cells. They can be used to neutralize the bioactivity of their cognate ligands in vitro and in vivo.

Synthesizing or amplifying the gene encoding the Fc-fused protein:

Free sequence optimization to meet your goals such as recombinant Fc-fusion protein expression at high and reliable levels. 

Construction, expression, and purification of the Fc-fusion protein:

* Fuse your protein of interest into human or mouse IgG Fc backbone (choice of human or mouse IgG1 Fc backbone; other IgG Fc backbone is available upon request) in mammalian expression vectors;
* Transiently express the recombinant Fc-fusion protein in mammalian expression systems, CHO or HEK 293 cells;
* Purify the recombinant Fc-fusion protein with Protein A resin and at least one additional protein polishing to ensure high quality: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <1 EU/mg (or <0.05 EU/mg if requested).

The Fc options include but are not limited to:
* Human IgG1-Fc, IgG2-Fc, IgG3-Fc, IgG4-Fc;
* Mouse IgG1-Fc, IgG2a-Fc, IgG2b-Fc, IgG3-Fc;
* Rat IgG1-Fc, IgG2a-Fc, IgG2b-Fc, IgG3-Fc;
* Rabbit IgG-Fc;
* Canine IgG-Fc.

Deliverable:
A. Purified protein from CHO transient production;
B. Study report.

BS050B:
Gene synthesis of the Fc fusion protein (additional fee);
Molecular construction of the recombinant Fc fusion protein;
0.1 Liter transient production of the recombinant Fc fusion protein in CHO cells (7-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <1 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050B-14:
Gene synthesis of the Fc fusion protein (additional fee);
Molecular construction of the recombinant Fc fusion protein;
0.1 Liter transient production of the recombinant Fc fusion protein in CHO cells (14-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <1 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050B-05:
Gene synthesis of the Fc fusion protein (additional fee);
Molecular construction of the recombinant Fc fusion protein;
0.1 Liter transient production of the recombinant Fc fusion protein in CHO cells (7-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <0.05 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050B-14-05:
Gene synthesis of the Fc fusion protein (additional fee);
Molecular construction of the recombinant Fc fusion protein;
0.1 Liter transient production of the recombinant Fc fusion protein in CHO cells (14-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <0.05 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050F:
DNA scale-up of the recombinant Fc fusion protein;
0.5 Liter transient production of the recombinant Fc fusion protein in CHO cells (7-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <1 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050G:
DNA scale-up of the recombinant Fc fusion protein;
1 Liter transient production of the recombinant Fc fusion protein in CHO cells (7-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <1 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050F-14-05:
DNA scale-up of the recombinant Fc fusion protein;
0.5 Liter transient production of the recombinant Fc fusion protein in CHO cells (14-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <0.05 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

BS050G-14-05:
DNA scale-up of the recombinant Fc fusion protein;
1 Liter transient production of the recombinant Fc fusion protein in CHO cells (14-day fed-batch cell culturing);
At least 2-step antibody protein purification;
Quality guarantee: SDS-PAGE >95%, SEC-HPLC >95%, and endotoxin <0.05 EU/mg. Mass spectrometry may be used to check molecular weight of the purified antibody.

Questions and Answers about the Recombinant Fc Fusion Protien Production Services:

Question: Why do you use at least 2-step protein purification methods to purify recombinant Fc fusion proteins?
Answer: Protein A or G affinity chromatography is commonly used for Fc fusion protein purification. However, for most Fc fusion proteins, one-step purification is not enough to remove impurity, such as aggregate of the proteins, endotoxin, and host cell protein or DNA residual. Impurity may interfere the in vivo and even in vitro activities of the purified Fc fusion proteins. The impurity may vary lot to lot, and result from many factors. To minimize lot to lot difference, it is better to obtain high purity Fc fusion proteins using multi-step purification methods.

Question: Is the yield from 14-day fed-batch cell culturing double that from 7-day cell culturing?
Answer: Generally speaking, yes. If not, it does not make much sense to do 14-day fed-batch cell culturing. Even double is not enough. Generally, we expect 2-10 folds of antibodies in the 14-day fed-batch cell culturing compared with the 7-day cell culturing.

Question: Our Fc fusion protein is a mouse IgG1. Are the prices the same as described above?
Answer: The cost of protein G purification is a little higher than protein A purification. The protein G resin is more expensive.

Question: We are doing high throughput screening (HTS) of a large number of recombinant Fc fusion proteins. Protein A one-step purification is enough. Can you do 30 ml, 10 ml or even 4 ml transient production?
Answer: Sure, please contact us.

We also provide the following protein production (DNA construction, protein expression and purification) services:
Albumin Fusion Protein Production Service
His Tag or FLAG Tag Fusion Protein Production Service

Applications:

ScFv-Fc fusion protein production: molecular construction of recombinant single chain antibodies (scFv-Fc), 0.1 liter transient production of recombinant scFv-Fc in CHO cells, and protein A purification of recombinant scFv-Fc

Recombinant IgG Fc Proteins:
Recombinant Human Fc Protein (IgG1)
Recombinant Human Fc Protein (IgG2)
Recombinant Human Fc Protein (IgG4)
Recombinant Mouse Fc Protein (IgG1)

Fc ELISA Kits and Reagents:
Human Fc ELISA Kit
Mouse Fc ELISA Kit
Human Fc ELISA Reagent Kit
Mouse Fc ELISA Reagent Kit