Do you lack the time and resources needed to scale up the production of your novel antibody? Let the antibody experts at ProteoGenix do it for you. Our recombinant antibody service is designed to provide you with 1 mg of your desired recombinant monoclonal antibody format in 4 weeks or less at an unbeatable price.Our proprietary mammalian expression system – XtenCHO™ – is the market’s leader in transient monoclonal antibody production. With the proven ability to perform proper antibody folding and glycosylation, XtenCHO™ outperforms even the most competitive commercial solutions like ExpiCHO™.Book a free consultation today to learn more.

Our process of recombinant
antibody production

Monoclonal antibody sequencing

  • Full-length or variable region sequencing

OR

Hybridoma sequencing

  • RNA isolation
  • cDNA synthesis
  • Amplification of antibody genes

OR

De novo antibody protein sequencing

  • Enzymatic digestion
  • LC-MS/MS separation
  • De novo peptide sequencing

Gene synthesis and cloning

  • Gene synthesis with codon optimization for the selected expression system
  • Subcloning into a high expression vector

Transfect

  • Plasmid amplification and preparation
  • Co-transfection of target cells

Transient expression

  • Small-scale expression of the antibody
  • Antibody purification and QC analysis

Advanced services

Analytics

Affinity (KD)
Stability
Aggregation

Purification
IEX
SEC
others

Stable Production
In CHO, HEK,
proprietary GS-…

Step Content Timeline Deliverables
Monoclonal antibody sequencing From hybridoma cell lines:
  • Hybridoma cell line provided by customer
  • RNA extraction and purification
  • Reverse transcription
  • cDNA amplification
  • Sequencing analysis
From purified antibody samples:
  • Antibody sample provided by customer
  • Enzymatic digestion of the antibody
  • LC-MS/MS separation
  • De novo peptide sequencing
  • Assembly of overlapping peptides
  • Data analysis
2 to 3 weeks for variable region & leader sequence
3 to 4 weeks for full length antibody
  • Detailed report
  • Antibody sequence
Gene synthesis and cloning
  • Gene synthesis
  • Codons optimization for Mammalian expression system
  • Subcloning into an expression vector
  • Test of two expression systems
  • Antigen production
2 to 3 weeks
  • Synthetic genes cloned into pUC57 plasmid
  • Certificate of analysis (CoA)
Monoclonal antibody production +purification
  • Plasmid amplification and preparation
  • Co-Transfection of target cells
  • Transient expression and purification
  • QC analysis (SDS-PAGE / UV280)
  • Antigen production
Starting from 3 to 4 weeks
  • Detailed report
  • Purified antibody

Options available (non exhaustive list):

  • Expression in alternative recombinant systems (bacterial, yeast, insect cells…)
  • Additional purification steps (IEX, SEC…)
  • Endotoxin assay and removal
  • Aggregate analysis : SEC-HPLC
  • KD determination : SPR (Biacore), SPRi (XelPlex), Octet Red96
  • Stability analysis : differential scanning calorimetry, IR

How much does our recombinant antibody production service cost?

The price of our recombinant monoclonal antibodies starts at a few hundred euros, making this an accessible option for many research and development projects. The primary factors influencing the cost are the number and amount of antibodies required. Generally, larger quantities or batches of antibodies will have a higher total cost, but the price per antibody may be reduced when ordering in larger volumes. This scaling effect can offer cost advantages for those seeking significant quantities of recombinant antibodies.

Additionally, the range of services selected by the customer can influence the overall cost. Besides the basic production of recombinant antibodies, customers may opt for additional services such as antibody analytics, purification, and stable production of the antibody in cell lines. Each of these supplementary services may contribute to the overall cost of the project.

It’s essential to keep in mind that while cost is a significant consideration, the quality, specificity, and functionality of the recombinant antibodies are equally crucial factors to evaluate when selecting a service provider. Investing in high-quality recombinant antibodies can lead to more reliable and reproducible results in research, diagnostics, and therapeutic applications.

How long does our recombinant antibody service take to complete?

The timeline for ProteoGenix’s recombinant antibody production service varies based on the intricacies of each project. Each project involves several essential steps, each contributing to the overall duration of the service. Here’s a general overview of the key stages and the approximate time frame:

1. Antibody Gene Isolation: This step entails hybridoma sequencing and de novo antibody protein sequencing to identify the target antibody gene. Depending on the complexity of the antibody, this stage can take around 1-2 weeks.

2. Gene Synthesis and Cloning: Once the antibody gene is identified, it needs to be synthesized and cloned into expression vectors. This process usually takes 1-2 weeks, depending on the gene length and any technical challenges.

3. Transfection and Expression: The recombinant antibody expression vectors are transfected into suitable host cells for antibody production. Small-scale expression is performed to verify antibody expression and functionality. This step can require 1-2 weeks.

4. Quality Control: After expression, the recombinant antibodies undergo rigorous quality control tests to ensure they meet the desired specifications. This can take around 1 week, depending on the number of antibodies and the extent of testing required.

Taking these stages into account, customers can typically expect to receive their recombinant antibody within 3-4 weeks from gene sequencing to final antibody production. However, the timeline may vary based on the complexity of the project, the number of antibodies, and any customizations or additional services you require.

Which Recombinant antibody formats can be produced with our service?

With our recombinant antibody service at ProteoGenix, we offer the production of a diverse range of monoclonal antibody formats that cater to various research and application needs. This includes modifying your antibody into one of the following antibody formats:

scFv (Single-chain variable fragment): scFv antibodies are engineered to combine the variable regions of the antibody’s heavy and light chains into a single polypeptide chain. These compact molecules retain antigen-binding specificity and are often used in research and diagnostic applications.

Fab (Fragment antigen-binding): Fab antibodies consist of the variable regions of the heavy and light chains, allowing them to retain antigen-binding capability. They also lack the Fc region, making them useful for certain applications where Fc-mediated effector functions are not required.

VHH (Nanobodies): VHH are single-domain antibodies derived from heavy-chain-only antibodies found in camelids. They are small and highly stable, making them valuable tools for research, diagnostics, and therapeutics.

Full-length Antibodies (IgG): Full-length antibodies, also known as IgG, are the conventional format used in many therapeutic applications. They possess intact Fc regions that facilitate effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC).

Antibody Fusion-Proteins: ProteoGenix also offers the production of antibody fusion proteins. These include antibodies fused with other functional proteins or tags, allowing for specific applications, such as affinity purification or targeted drug delivery.

Our commitment to offering a diverse selection of antibody formats ensures that you find the most suitable option to address your specific scientific and medical requirements.

The choice of format depends on your intended application. Our team of experts is always available to provide guidance and assistance in selecting the optimal recombinant monoclonal antibody format for each unique project.

What are recombinant monoclonal antibodies?

Recombinant antibodies are synthetic antibodies produced using recombinant antibody technology. Unlike traditional antibodies derived from immune responses in animals, recombinant antibodies are designed and engineered in the laboratory. This process involves isolating the genes that encode the antibody’s variable regions and then expressing these genes in host cells to generate the desired antibody.

Production Process:
The recombinant antibody production process typically involves several key steps. First, the genes encoding the antibody’s variable regions are isolated, either from hybridoma cells (for known antibodies) or through de novo sequencing. Next, these genes are synthesized or cloned into expression vectors. These expression vectors are then transfected into suitable host cells, such as mammalian cells or bacteria, to produce recombinant antibodies.

Characteristics:
Recombinant antibodies offer several advantages over traditional antibodies. Since they are engineered, their binding affinity and specificity can be optimized for specific targets. This allows for the creation of high-affinity antibodies with reduced cross-reactivity. Recombinant monoclonal antibodies can also be designed to lack the Fc region, eliminating potential Fc-mediated effector functions, and making them suitable for certain therapeutic and research applications.

Consistency:
Recombinant antibody production ensures a high degree of consistency and reproducibility. The genetic engineering process allows for precise control over the antibody’s sequence, leading to a more uniform product with predictable characteristics. This consistency is crucial, especially in therapeutic applications where precise dosing and efficacy are essential.

Scalability:
Another significant advantage of recombinant antibodies is their scalability. Once the gene sequences are available and the production process is established, it becomes relatively straightforward to scale up production to generate large quantities of the antibody. This scalability is vital in meeting the demand for research, diagnostics, and therapeutic applications.

Recombinant antibody production has become an essential tool in research, diagnostic, and therapeutic applications. Their ability to be precisely engineered for specific targets, consistent quality, antibody format, and scalability makes them valuable assets in addressing a wide range of scientific and medical challenges. As technology and techniques in genetic engineering continue to advance, the potential applications and impact of recombinant antibodies are expected to grow significantly in the future.

 

XtenCHO™: The gold standard expression system for transient monoclonal antibody production

Looking for fast antibody production at competitive prices? XtenCHO™ makes it possible.

Thanks to our in-house Xten transfection protocol, proprietary cell line XtenCHO™, 28+ years of experience in recombinant production, and over 500 successful antibody production projects, we have the market’s most competitive solution.

With production yields consistently exceeding even those of ExpiCHO™, we are equipped to overcome your unique antibody expression challenges while keeping production costs low.

Here are some examples of the incredible results obtained in the transient expression of biosimilars

For more detailed results about the use of XtenCHO™ for recombinant antibody production, discover other unbelievable results by downloading our poster or watching Dr. Marcelo Viegas presentation made at the Festival of Biologics (2020).

Case study: Pembrolizumab recombinant antibody production

Aim Of The Study

Here, we showcase the recombinant monoclonal antibody production of Pembrolizumab using our proprietary Xten expression system.

Genes And Expression Vectors

The genes coding for the Heavy chain (HC) and the light chain (LC) of the therapeutic antibody Pembrolizumab were chemically synthesized with optimizations for expression in CHO cells. The genes were cloned in ProteoGenix’s proprietary mammalian cells expression vector pTXs1. The vector map is provided in the PDF report.

Small-scale Recombinant Monoclonal Antibody Production And Purification Tests

An endotoxin-free DNA preparation was done for the pTXs1 expression constructions. By using the proprietary Xten transfection protocol, the plasmids were then transiently transfected in the proprietary XtenCHOTM cells.

Culture medium was collected and production yield was determined by Octet Red analysis. The recombinant antibody was then purified by affinity vs. protein G.

Recombinant antibody production QC

QC of the purified Pembrolizumab. Coomassie blue staining.
A. Non-reduced SDS-PAGE. B. Reduced SDS-PAGE.

Antibody Production Yield Purity
Pembrolizumab 235mg/L >95%

Recombinant Antibody Service Benefits

Recombinant antibody production has been a major breakthrough in the therapeutic antibody development context as it leads to several advantages over “traditional” antibody production in hybridoma cell lines.

1. Optimal reproducibility, control and security

Monoclonal antibody production for therapeutic applications requires high control and reproducibility. Because crucial parameters such as DNA and protein sequence are chemically defined, recombinant antibody production allows creating a standardized process for monoclonal antibody production with very low batch-to-batch variations. As recombinant antibody production starts from a known antibody or DNA sequence, mutations or cell-line drift that can occur in hybridoma cell lines are also obviated.

2. High purity

Recombinant antibody production leads to high purity grade due to the absence of serum contaminants.

3. Ease of engineering and reformatting

Recombinant monoclonal antibody production allows getting access to the control of the antibody structure at the genetic level. This is particularly important when it comes to therapeutic antibody development as it permits modulatingmodulating the pharmacological properties of the recombinant antibody.

The possibility to modify the gene that will further be expressed by a selected expression system also gives access to an infinite range of solutions in terms of:

  1. Reformatting: species, isotype or subtype switching
  2. Antibody engineering: monoclonal antibody humanization, affinity maturation, bispecific antibody, antibody drug conjugates (ADC), antibody fragments (Fab, Fab2, scFv).

4. No animal use

ProteoGenix is dedicated to the highest standards of medical ethics and animal welfare. Thanks to recombinant antibody production technology, the antibodies can be quickly produced without relying on animal use in any step of the process.

Transient or stable? Which recombinant antibody
production service should you choose?

In this section, we will highlight the difference between transient and stable antibody expression.

Recombinant antibody production offers a main advantage: it allows rapid production of a relatively large number of antibodies. However, in transient transfection, the transfected gene does not integrate the host genome and can be lost in several days (over a short time period, of several days). Thus, it can be the perfect solution for short-term applications such as small-scale protein expression.

3 months), stable cell lines are privileged for long-term applications such as therapeutic antibody production. For large-scale bioproduction, a stable cell line allows to construct of a standardized process that guarantees higher yield and quality control.

For any additional question, please feel free to contact our PhD account manager who will be glad to guide you toward the most suitable antibody production solution.

Cell Line Characteristic Stable Cell Line Expression Transient Transfection
Duration of Expression Long-term Short-term
Expression Level High Moderate to High
Stability Stable Expression Temporary Expression
Time and Effort ++ +
Upfront Investment ++ +
Selection and Screening Required Not required
Protein Quality Control Consistent Less Consistent
Protein Modification Limited Possible, but more complex
Scalability Excellent Limited to batch sizes
Application Suitability Long-term assays, stable Rapid screening and initial

For any additional question, please feel free to contact our PhD account manager who will be glad to guide you toward the most suitable antibody production solution.

What are the applications of transient antibody expression?

Transient antibody expression in mammalian systems is useful for many applications. These systems combine proper protein folding and post-translational modification with short lead times making them invaluable for:

  • Screening of drug candidates: : transient expression allows the production of defined quantities of antibodies in systems very similar to those used for commercial biopharmaceutical production. In this way, transient production allows a preliminary characterization of these molecules as well as the detection of potential production hurdles exponentially increasing antibody developability.
  • Antibody variant screening: antibody variants are frequently produced in an attempt to improve certain biophysical properties of antibodies including affinity and stability. Variants are also produced during the process of antibody humanization. In this case, the small-scale production of several variants helps selecting the best candidates for further development.
  • Small scale production for analytical applications: these systems allow the quick production of antibodies or fragments thereof in many different formats. Given that mammalian systems efficiently secrete these molecules to the culture media, they can be purified using inexpensive methods such as protein A/G purification. These highly pure antibody batches can be easily conjugated with enzymatic or fluorescent tags for research or diagnostic applications.

Recombinant antibody production: cells and approaches

Mammalian expression systems based on Chinese hamster ovary (CHO) or Human embryonic kidney (HEK) cells are the industry’s most robust solutions for recombinant antibody expression.

Their ability to perform human-like glycosylation (crucial to reduce immunogenicity of antibodies), proper protein folding and their efficient secretory pathways, makes the process ideal for therapeutic applications. Cell used for antibody production must be able to maintain a high productivity and batch-to-batch consistency. All these criteria are met by our proprietary cell line XtenCHO™.

Our vast expertise in producing antibodies in CHO and HEK systems is also complemented by our 28+ years of experience in protein production in a number of systems including bacterial, yeast, and insect cells.

The use of these systems for recombinant antibody expression continues to grow in popularity due to the growing demand for solutions with lower costs and shorter lead times. Although XtenCHO™ remains our gold standard for antibody production, we are also able to produce full-length or antibody fragments in all these alternative systems.

What are the advantages and limitations of these alternatives in comparison to mammalian production systems?

Mammalian systems Bacterial systems Yeast systems
Advantages
  • Proper protein folding
  • Human-like glycosylation
  • Extracellular production – easier purification
  • High batch-to-batch consistency
  • High purity
  • High production yields meeting important quality standards
  • Adapted to any antibody format including full-length, antibody fragments, or antibody conjugates…
  • High production yields
  • Shorter lead times (in comparison to mammalian cells)
  • Advantageous for antibody fragment production (scFv, Fab, VHH)
  • Ease of genetic manipulation
  • No stable integration required for large-scale production
  • Ease of scaling up production
  • Extracellular production – easier purification
  • Ease of genetic manipulation
  • High production yield
  • Shorter lead times (in comparison to mammalian cells)
  • Post-translational modifications (but not human-like glycosylation)
  • No stable integration required for large-scale production
  • Ease of scaling up production
Limitations
  • Longer lead times
  • Time-consuming and laborious when creating stable cell lines
  • Cell lines harder to engineer
  • No post-translational modifications
  • Intracellular production – harsher purification methods required
  • Risks of forming inclusion bodies
  • Contamination with endotoxins
  • Moderate risk of plasmid loss even in selective conditions
  • Inability to perform human-like glycosylation without extensive engineering

    Which system should you choose for the recombinant expression of your antibody?

    Each project is evaluated on a case by case basis thus we are always able to propose the best solution according to the structure of your antibody, intended application, available budget and timeline, and scale of production.

    Learn more about our protein expression services in:

    Bacteria

    Discover our protein expression service in Escherichia coli or Bacillus subtilis and benefit from short lead time and high productivity

    Yeast

    Learn more about our protein production protocols in Saccharomyces cerevisiae or Pichia pastoris for optimal production and process scalability

    Insect cells

    Produce your protein leads in baculovirus/insect cells for improved protein folding and accurate post-translational modifications

    Monoclonal antibody production
    from gene to bioproduction

    The successful production of recombinant antibodies starts with a careful optimization of genes, vectors, and transfection protocols. These apparently inconsequential details represent one of the most important antibody production steps of the process. All these parameters must be compatible with the requirements of the cells used for antibody production.

    Access our page dedicated to the most frequently asked questions (FAQs) about recombinant antibodies, to learn more about all stages of the production process as well as ProteoGenix’s recommendations to help you achieve high and consistent levels of production in your project.

    ProteoGenix has guided thousands of researchers throughout their monoclonal antibody production projects since 2003. As a one-stop solution provider, we master the whole antibody development process from antibody generation by hybridoma development or antibody phage display to recombinant antibody production in XtenCHOTM and stable cell line generation. Trusting ProteoGenix, a monoclonal antibody production leader, is a guarantee to take your project to next level!