Fermentation for Recombinant Proteins
As a leading CDMO, BOC Sciences provides one-stop services to manufacture recombinant proteins. With advanced fermentation and purification technologies, our team of leading scientists supports the development and large-scale production of recombinant proteinsto meet diverse needs in scientific research, pharmaceuticals, agriculture, and more, accelerating the progress of your projects.
What is Recombinant Protein?
Recombinant protein is a protein encoded by recombinant DNA. Modification of the gene can lead to expression of a mutant protein. The use of recombinant DNA technology to express recombinant proteins in unnaturally occurring cells is known as heterologous protein production. Recombinant DNA technology has made possible the large scale production of proteins that were previously unavailable, relatively expensive or difficult to obtain in large quantities. These recombinant proteins have broad applications in vaccine development, biopharmaceuticals, diagnostic reagents, and biological research. Recombinant proteins are formed by transfecting foreign genes into a host cell, and the steps for producing recombinant proteins include:
- Isolate gene of interest
- Insert gene of interest into plasmid DNA
- Transform recombinant DNA plasmid into host
- Plate cells onto agarose gels
- Expand the positive clones to exponentially replicate copies of desired DNA
- Expression of proteins of interest
Host of recombinant proteins
The strains commonly used to produce recombinant proteins include Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris, CHO cells and baculovirus-infected insect cells. E. coli is widely used as a host for heterogeneous protein expression because of its ease of growth and manipulation, its well-known knowledge of genetics and biology, its ability to achieve expression quickly, its proven fermentation technology. In contrast to E. coli, which is unable to perform eukaryotic-specific post-translational modifications due to its status as a prokaryote, S. cerevisiae and P. pastoris, as eukaryotic microorganisms, are able to produce recombinant therapeutic proteins closer to those of humans. CHO cells are one of the most popular hosts for recombinant proteins, used for the production of monoclonal antibodies and more humanlike therapeutic proteins. The baculovirus-insect cell system also supports the production of a large number of proteins with complex post-translational modifications.
Category of recombinant proteins
Recombinant proteins are widely used in the pharmaceutical industry, such as antibodies or antibody fragments and enzymes for disease treatment, protein-based polymers for drug delivery, and recombinant protein scaffolds for tissue engineering, etc.
| Types of recombinant proteins | Examples | |
|---|---|---|
| 1 | Industrial enzymes | Amylases, lipase, chymosin, glucose oxidase, pullulanase, enterokinase, invertase, cellulase, xylanase |
| 2 | Therapeutic proteins | Insulin,insulin glargine, insulin lispro, Fab and ScFv fragments, monoclonal antibodies, recombinant vaccines, interferons |
| 3 | Therapeutic enzymes | Streptokinase, urokinase, trypsin, glutaminase, B-lactamase, l-asparaginase, glucosidase, collagenase, uricase |
| 4 | Protein scaffolds | Human collagen, bacterial collagen, spider silk analogs, coiled-coil silk, tropoelastin, elastin segments, resilin |
Our Services
- Strain improvement, such as genetic recombination, cloning, transfection, etc.
- Fermentation upstream and downstream process optimization, such as the use of single-use systems.
- Fed-batch or continuous fermentation in high-cell-density.
- Purification process development, such as membrane chromatography.
- One-stop services for recombinant proteins from host strains development to commercial scale-up manufacturing.
Our Fermentation Capabilities
- Lab-scale: equipped with 30 L to 200 L fermentation tanks
- Pilot-scale: equipped with fully automatic fermentation tanks of different volumes ranging from 4,000 L to 12,000 L
- Commercial-scale: Fermentation tanks of 10 to 120 m3
Our Advantages
- Consistent process and quality product development using QbD (Quality-by-Design)
- Efficient upstream and downstream process development
- High throughput equipment and GMP facilities
- Flexible, automated fermentation platform and economical product development
- High product yield and quality and low cost
- Investment in environmental protection measures and skills
- Well-established quality system, which has passed through a series of official inspections and customer audits
Application of recombinant proteins
(1) Biopharmaceuticals
Recombinant proteins play a crucial role in the biopharmaceutical industry, including monoclonal antibodies and recombinant protein drugs. Our fermentation-based recombinant protein services provide high-quality protein materials to accelerate drug research, development, and production.
(2) Vaccine Development
Recombinant protein vaccines are a new generation of vaccines offering high safety and strong immunogenicity. Our services provide recombinant protein antigens to support rapid vaccine development and commercialization.
(3) Diagnostic Reagents
Recombinant proteins serve as essential raw materials for antigens or antibodies in diagnostic reagents, ensuring high purity and high activity. We offer stable protein supplies to diagnostic reagent companies, ensuring product quality and accuracy.
(4) Biological Research
Recombinant proteins are indispensable tools for studying protein functions, structures, and interactions. Our services provide high-quality recombinant proteins to support cutting-edge biological research.
Fermentation Recombinant Protein Service
Custom Recombinant Protein Expression
Gene Cloning and Sequence Optimization: Optimize the provided target gene sequence to ensure efficient expression.
Expression Vector Construction: Choose the appropriate expression vector, such as E. coli, yeast, or mammalian cell vectors, to meet the expression needs of different proteins.
Host Cell Selection and Transformation: Provide various host cell systems (such as E. coli, yeast, insect cells, CHO cells, etc.) and complete efficient transformation.
Fermentation Process Development and Optimization
Lab-Scale Fermentation: Conduct small-scale fermentation in the lab to optimize fermentation conditions and ensure efficient protein expression.
Pilot-Scale Amplification: Offer fermentation scale-up services from lab to pilot scale, addressing technical bottlenecks in large-scale production.
Large-Scale Production: Support industrial-grade fermentation production to meet customer demands for large quantities of recombinant proteins.
Protein Purification and Modification
Purification Process Development: Design an efficient purification process based on the protein characteristics to ensure product purity.
Protein Modification Services: Provide customized services such as tag removal, glycosylation modification, phosphorylation modification, etc., to meet special application requirements.
Quality Testing and Analysis
Purity Analysis: Use techniques such as HPLC and SDS-PAGE to test protein purity.
Activity Testing: Assess protein function through bioactivity assays (such as ELISA, cell experiments).
Safety Testing: Test endotoxins, host protein residues, and other indicators to ensure product safety.
One-Stop Solution
From Gene to Protein: Provide a one-stop service from gene sequence to recombinant protein product, so customers do not need to build complex technical platforms themselves.
Customized Service: Offer personalized solutions based on customer needs to meet different application scenarios and functional requirements.
Our Capability
We have high end production and processing equipment, including fermentation tanks of different sizes, continuous sterilization, multiple aseptic feed lines, and separation and purification equipment.
- Lab-scale: equipped with 5L to 200L fermentation tanks imported or domestic source.
- Pilot-scale and Commercial-scale: equipped with fully automatic fermentation tanks of different volumes to 12,000L.
- Commercial scale: Fermentation tanks of 10-120 m3
- 4 Manufacturing sites
- 40+ reactors of from 30-4000 L, 170+ reactors from 20 KL to 30 KL, 24+ reactors of >100 KL
- 2 Hydrogenation reactors (200 L, 4Mpa and 1000L, 4Mpa)
Our Advantages
- Consistent process and quality product development using QbD (Quality-by-Design)
- Efficient upstream and downstream process development
- High throughput equipment and GMP facilities
- Flexible, automated fermentation platform and economical product development
- High product yield and quality and low cost
- Investment in environmental protection measures and skills
- Well-established quality system, which has passed through a series of official inspections and customer audits
- Fast growth and high-density fermentation-Microbial hosts like E. coli and yeast grow rapidly and achieve high cell densities, significantly increasing recombinant protein yield.
- Short production cycle-Fermentation technology enables large-scale production in a relatively short time, meeting industrial demands.
Fermentation Recombinant Protein Service Process
1. Project Consultation and Requirement Analysis
Customers can contact us via online customer service, email, or phone to provide project requirements and target protein information. Our technical team will have detailed communication with the customer to understand the characteristics, application scenarios, and special requirements of the target protein. Based on customer needs, we will provide technical solution suggestions.
2. Quotation and Contract Signing
Provide a detailed quotation and service terms based on project requirements. After both parties confirm the cooperation details, a service contract will be signed, specifying the project scope, delivery standards, and time milestones.
3. Project Start and Technical Development
Gene Cloning and Vector Construction: Synthesize genes, clone them, and construct expression vectors based on the gene sequence provided by the customer.
Host Cell Selection and Transformation: Choose the appropriate host cell system and complete the transformation and screening of the target gene.
Small-Scale Fermentation and Optimization: Conduct small-scale fermentation in the laboratory, optimizing fermentation conditions to ensure efficient protein expression.
4. Pilot-Scale Amplification and Large-Scale Production
Scale up the optimized fermentation process from the small-scale to pilot scale, addressing technical issues in large-scale production. Perform industrial-grade fermentation production based on customer needs, ensuring stable protein yield and quality.
5. Protein Purification and Quality Testing
Design an efficient purification process based on protein characteristics to ensure product purity. Use techniques such as HPLC, SDS-PAGE, and ELISA to test protein purity, activity, and safety indicators. Provide a detailed testing report to ensure customers have a clear understanding of the product quality.
6. Delivery
Deliver the qualified recombinant protein product to the customer according to the contract.
FAQ
1. What host cell systems do you support?
We offer a variety of host cell systems, including Escherichia coli, yeast (such as Pichia pastoris and Saccharomyces cerevisiae), insect cells (Baculovirus expression system), and mammalian cells (such as CHO cells). We select the most suitable expression system based on the characteristics of the target protein and customer requirements.
2. What quality testing services do you provide?
We provide comprehensive quality testing services, including:
Purity Testing: Protein purity is detected using techniques such as HPLC, SDS-PAGE, etc.
Activity Testing: Protein function is assessed through bioactivity experiments, such as ELISA and cell assays.
Safety Testing: We test for endotoxins, host protein residues, and other indicators to ensure product safety.
3. Do you offer customized services?
Yes, we offer fully customized services. Based on customer needs, we can optimize expression vectors, adjust fermentation conditions, and perform protein modifications (such as tag removal, glycosylation modifications), among other services, to meet specific application requirements.
