Question 1

Can your platform handle complex, unstable, or low-immunogenic targets?

Accepted Answer

Yes. Our phage display platform is designed to handle challenging targets, including complex, unstable, toxic, or low-immunogenic antigens. With more than 500 successful phage display projects, we routinely deliver high-quality binders even when targets are difficult to express or highly conformational.

Question 2

What antigen formats are compatible with the panning workflow (soluble, cell-based, peptides, membrane proteins…)?

Accepted Answer

We can work with a wide range of antigen formats, including peptides, proteins, small molecules, and whole cells. For membrane proteins and clinical targets where native conformation matters, we recommend cell-based panning to select binders that recognize the target in a physiologically relevant form.

Question 3

Do you offer specialized panning strategies such as negative selection or competitive elution?

Accepted Answer

We’ve developed an optimized process for phage display to perform negative selection and/or competitive elution to ensure you obtain the antibodies with the ideal specificity and/or activity like a neutralizing activity.

Question 4

What assays do you use for hit screening and affinity assessment?

Accepted Answer

Primary hit identification is based on ELISA screening to confirm binding and select high-affinity candidates. If your application requires a specific validation format, we can further screen individual clones by ELISA, Western blot, or flow cytometry on whole cells. We also offer affinity (KD) measurement as a downstream characterization step.

Question 5

Can you deliver both full-length IgGs and fragments (scFv, Fab, VHH) depending on the project?

Accepted Answer

Our libraries and screening services are available in scFv, Fab, and VHH formats to match your application. After hit discovery, we can clone your sequences into expression vectors and produce recombinant antibodies in the format best suited to your next steps.

Question 6

Do you provide sequence information for selected clones?

Accepted Answer

Yes. Your deliverables include a complete report with the DNA and amino-acid sequences of selected clones. You also receive full rights to the sequences so you can freely develop, engineer, or manufacture your antibodies.

Question 7

Can I request a customized panning strategy for my specific application?

Accepted Answer

Your dedicated PhD-level account manager will be happy to discuss about specific panning strategy you would like to use and advise on the best possible option to increase your chances of success.

Question 8

Do you perform counter-selection to reduce cross-reactivity?

Accepted Answer

Upon customer request, we can perform counter-selection and use a dedicated phage display strategy to enrich the number of candidates binding to the protein of interest and not to proteins which should not be recognized.

Question 9

How do you ensure that selected binders are specific, high-affinity, and physiologically relevant?

Accepted Answer

We combine multiple rounds of biopanning with ELISA-based screening to enrich and confirm high-affinity, specific binders. When relevance to native biology is critical, for example with membrane proteins, we use whole-cell panning to keep the target in its natural conformation and select antibodies that perform in real conditions.

Question 10

How long does the full phage display campaign typically take?

Accepted Answer

Thanks to our pre-built libraries and optimized workflow, you can receive validated antibodies in as little as 4 weeks.

Question 11

Are your libraries proprietary, and what advantages do they offer in terms of diversity and success rate?

Accepted Answer

Yes. Our phage display libraries are proprietary and exclusive to ProteoGenix, giving you access to resources no other provider can offer. They combine extensive diversity across species and formats with exceptional quality, including an in-frame frequency above 93 percent, which translates into a higher probability of strong, developable hits.

Question 12

What quality controls are performed before delivering the selected clones or purified antibodies?

Accepted Answer

SDS-PAGE and concentration determination by UV280/Bradford are performed in standard, but we can perform many other QCs on purified antibodies, such as : ELISA, SPR/BLI, SEC-HPLC, nanoDSF, MS, endotoxin assay&#8230;

Question 13

Can your team support downstream steps such as affinity maturation, humanization, or reformatting?

Accepted Answer

Yes. Beyond screening, we can support every next step, including affinity maturation, humanization, and reformatting into advanced formats such as bispecific antibodies or ADCs. We also provide cloning, recombinant production, and in-depth characterization to accelerate your program.

Question 14

Are your antibodies suitable for diagnostic, therapeutic, or research applications?

Accepted Answer

Yes. Our phage display services are built to support research, diagnostic, and can be part of our therapeutic antibody discovery services. We tailor the library choice and validation strategy to your application so you get binders that perform in your conditions.

Question 15

What types of packages or project scopes are available (full discovery, screening only, library access…)?

Accepted Answer

Our phage display services are fully customizable. You can rely on us for the full discovery journey, from library selection to validated binders, or request screening-only support for a specific step. Every project gives you access to our complete portfolio of proprietary libraries. If you prefer to run screening internally, you can also purchase a library with full usage freedom and no hidden fees.

Question 16

Do you offer scale-up or recombinant production for selected clones?

Accepted Answer

Yes. After selection, we can subclone your antibody cDNAs into expression vectors and produce recombinant antibodies in the format and scale required for your workflow. We can guide you from high-throughput expression and screening to large-scale transient expression and stable cell line development.

Question 17

What post-project support do you provide (additional screening, production, long-term storage…)?

Accepted Answer

We can extend screening to confirm performance in your specific assay, produce recombinant antibodies, engineer next-generation formats, and run advanced characterization such as affinity or stability testing. This allows you to carry out your entire custom antibody development with a single partner.
We can also provide additional services such as long-term storage. For a one-time fee, ProteoGenix can manage storage for 1 year or indefinitly.

Question 18

What Is Hybridoma Technology and Its Significance?

Accepted Answer

Hybridoma technology is the cornerstone of our custom hybridoma monoclonal antibody services, creating cell lines that produce monoclonal antibodies with unmatched specificity. This method is essential for advancing research, diagnostics, and therapeutic solutions, offering antibodies with superior stability and affinity.

Question 19

Which Antigens Are Suitable for Hybridoma Development at ProteoGenix?

Accepted Answer

Our hybridoma antibody production services are versatile, accommodating a wide range of antigens, from peptides to eukaryotic cells. Through peptide synthesis and DNA immunization, we tailor our approach to each project, ensuring optimal antigen presentation and immune response.

Question 20

How Does ProteoGenix Guarantee High-Quality Monoclonal Antibodies?

Accepted Answer

Quality is paramount in our hybridoma monoclonal antibody production services. We begin with a strategic immunization plan, using either in situ DNA or synthetic peptides as immunogens, to maintain antigen integrity and stimulate a targeted immune response, ensuring the production of high-affinity, biologically relevant antibodies.

Question 21

Can ProteoGenix Produce Antibodies for Both Therapeutic, Research and Diagnostic Uses?

Accepted Answer

Yes, our custom hybridoma development process yields antibodies suitable for a broad spectrum of applications. From diagnostic assays requiring high sensitivity to therapeutic interventions needing humanized antibodies as well as research tools critical for academic publications, we leverage our expertise to meet diverse application demands.

Question 22

What Distinguishes ProteoGenix's Hybridoma Development Services?

Accepted Answer

Our hybridoma production services stand out due to over 20 years of experience and a success rate exceeding 90%. We excel in managing complex projects, from antigen design to antibody production, ensuring each antibody meets our high standards of affinity and specificity.

Question 23

What Is the Timeline for Hybridoma Development at ProteoGenix?

Accepted Answer

The duration of our custom hybridoma generation process varies, but we aim for efficiency with positive clones available within 45 days, thanks to our fast-track protocol. The entire project typically spans 90-120 days, including time for thorough validation testing.

Question 24

What Antibody Yields Can Be Expected from ProteoGenix's Hybridoma Services?

Accepted Answer

Our hybridoma development services yield antibodies ranging from 40 mg/l in serum-containing mediums to 20 mg/l in serum-free conditions. We guarantee antibodies of the highest purity (>95%), ensuring they&#8217;re suitable for various applications, including in vivo use.

Question 25

How Are Biologically Relevant Antibodies Developed?

Accepted Answer

We prioritize antigen design to ensure the development of biologically relevant antibodies. Our comprehensive evaluation phase allows for tailored immunization strategies and screening processes, ensuring the antibodies produced are perfectly aligned with your project&#8217;s goals.

Question 26

How Are Hybridoma Cell Lines Screened for Antibody Activity?

Accepted Answer

Initial screening employs ELISA, complemented by flow cytometry and other assays to ensure antibodies meet application-specific requirements. This meticulous screening process guarantees our antibodies&#8217; optimal performance across diagnostic and therapeutic applications.

Question 27

What are the advantages of using hybridomas?

Accepted Answer

Hybridoma technology offers several key benefits:

Unlimited antibody production: Hybridomas are theoretically immortal, allowing for long-term antibody production. However, their &#8220;immortality&#8221; can be affected by genetic drift and mutations, which may compromise stability and productivity over time. To ensure you never lose access to your antibody, we recommend using our antibody sequencing services. This is the most reliable solution to guarantee continuous antibody production in the long run. And if you no longer have access to the hybridoma itself, don’t worry—we can still retrieve your sequence from a purified antibody sample through our de novo antibody sequencing service.
Naturally high-affinity antibodies: Hybridomas are derived from B cells that have undergone in vivo affinity maturation, resulting in antibodies with naturally high affinity and specificity.
Antibodies with high developability: This method preserves the natural pairing of heavy and light variable chains (VH/VL pairing), which enhances antibody developability and ensures optimal performance for downstream applications.

Question 28

What Is a Hybridoma?

Accepted Answer

Hybridomas are immortalized cell lines, key to our custom hybridoma development services, that produce monoclonal antibodies. Created by fusing B cells with myeloma cells, this method, pioneered by Georges Kohler and Cesar Milstein in 1975, utilizes electrofusion or polyethylene glycol (PEG) for the continuous production of high-quality antibodies.

Question 29

What Are the Main Sources for Hybridoma Creation?

Accepted Answer

Rodents, such as mice, rats, and hamsters, are the primary sources for our hybridoma monoclonal antibody production services due to their effective monohybridoma production. We also explore heterohybridomas from different species, including chickens, rabbits, or humans, to expand our antibody generation capabilities, despite their potential instability.

Question 30

How Are Hybridomas Grown In Vitro?

Accepted Answer

Our hybridoma production services cultivate hybridomas in suspension cultures, a humane method aligned with serum-free and chemically-defined mediums. This approach minimizes contamination and supports the production of high-purity antibody stocks, reflecting our commitment to ethical and efficient antibody production.

Question 31

What Are the Principal Uses of Hybridoma Cell Lines?

Accepted Answer

Hybridoma cell lines, central to our custom hybridoma generation process, are crucial for producing IgG monoclonal antibodies for diagnostics and research. These antibodies can be modified for specific antigen detection, and through hybridoma sequencing, we ensure their long-term productivity and adaptability for recombinant expression.

Question 32

Why Is Hybridoma Development Vital for Antibody Production?

Accepted Answer

Hybridoma technology, foundational to our hybridoma antibody production services, enables the unlimited and reproducible creation of monoclonal antibodies. This cost-effective method is essential for generating antibodies with preserved natural heavy and light chain pairing, offering high stability and specificity for diagnostics and therapeutics.

Question 33

Ensuring the Production of Highly Specific Antibodies via Hybridomas?

Accepted Answer

To guarantee the specificity of antibodies, our hybridoma development services employ strategic immunization techniques. By preserving the antigen&#8217;s native structure through DNA immunization or recombinant expression, we ensure the generation of antibodies tailored for membrane-bound targets or specific epitopes, crucial for their intended diagnostic or therapeutic applications.

Question 34

How Does B Cell Activation Affect Hybridoma Performance?

Accepted Answer

B cell activation is pivotal in our custom hybridoma development services for producing monoclonal antibodies. Mature B cells, activated in a T cell-dependent manner, are essential for generating antibodies with high specificity and affinity, significantly enhancing the quality and performance of hybridoma-produced antibodies.

Question 35

What Is T Cell-Dependent Activation and Its Role?

Accepted Answer

T cell-dependent activation, a cornerstone of our hybridoma monoclonal antibody production services, involves the presentation of antigens to B cells alongside helper T cell involvement. This process, crucial for a strong immune response, fosters the production of high-affinity antibodies, underpinning successful hybridoma development.

Question 36

Which Applications Are Most Suited for Hybridoma-Generated Antibodies?

Accepted Answer

Hybridoma-generated antibodies, central to our custom hybridoma generation process, excel in diagnostics, therapeutics, and research. Their high specificity and stability make them ideal for precise antigen detection in ELISA and flow cytometry, and they are pivotal in developing therapeutic antibodies for complex diseases.

Question 37

Can One Hybridoma-Generated Antibody Fit All Diagnostic Needs?

Accepted Answer

While our hybridoma antibody production services yield highly specific antibodies, a single antibody may not meet all diagnostic criteria. The suitability of an antibody for a particular diagnostic application depends on its affinity, antigen nature, and assay conditions, necessitating tailored antibody selection.

Question 38

What Are Other Applications for Hybridoma-Generated Antibodies?

Accepted Answer

Beyond their primary use in diagnostics and therapeutics, antibodies produced through our hybridoma production services find roles in basic research, protein interaction studies, and vaccine development. They&#8217;re also instrumental in bioengineering for antibody-drug conjugates and in agriculture for enhancing disease resistance.

Question 39

How Much Does Single B Cell Sorting Cost?

Accepted Answer

ProteoGenix’s B cell sorting services offer a flexible and customizable pricing structure to cater to your individual needs. Our prices start at a few thousand euros with pricing depending on the number of specific requirements for each project. The pricing is largely influenced by the number of antibody sequences to be analyzed and the scope of the service purchased.
We also offer the option for customers to enhance their single B cell sorting experience with supplementary services. These additional services include ELISA for positive/negative screening against specific peptides, proteins, or small molecules, as well as cross-reactivity profiling. Furthermore, our Competitive ELISA service enables the identification of clones with blocking or neutralizing activity, providing invaluable insights for therapeutic development.
Building a custom antibody using B cell isolation is a valuable investment that yields long-term cost savings. By precisely selecting high-affinity antibody clones early in the development process, it minimizes resources spent on less promising candidates. The targeted approach reduces downstream costs, ensuring efficient large-scale production and accelerating the path to a successful therapeutic, resulting in substantial savings in the long run.
At ProteoGenix, we understand the uniqueness of each project and aim to provide tailored solutions for our clients. Our team of experts works closely with customers to evaluate their specific needs and propose the most appropriate services within their budget. With our transparent and ranking-based pricing approach, customers can confidently choose the services that best align with their research objectives, ultimately ensuring the success of their single B cell sorting projects.

Question 40

How Long Does Single B Cell Isolation Take?

Accepted Answer

At ProteoGenix, our single B cell antibody technologies are designed for rapid and efficient results. The customer can get his antibody within only 6 weeks, ensuring a quick turnaround for their project.
We are proud to introduce our new immunization protocol that offers significant time savings, allowing us to complete the process 50% faster than our competitors. Immunization delays have been reduced from the typical 6-8 weeks to just 4-6 weeks, further streamlining the B cell sorting timeline and providing customers with even faster access to their desired antibody sequences.

Question 41

Can I patent my antibody sequence after B cell sorting?

Accepted Answer

You can patent your antibody sequence that is delivered from our B cell sorting service. We believe in providing our customers with full ownership of their antibody sequences, giving you the freedom and protection to secure intellectual property rights for your valuable discoveries.
With our B cell sorting services, you can confidently pursue patent applications and protect the innovative potential of your custom antibodies, ensuring their exclusivity and commercial value in the market.

Question 42

Is a Single B-cell sorting optimal for challenging antibody targets?

Accepted Answer

Traditional monoclonal antibody technology involves creating a hybrid cell line by fusing B-cells from immunized mice with an immortal B-cell myeloma cell line. The resulting fused cells are referred to as hybridoma cells because they are a hybrid between a healthy B-cell and a transformed B-cell. Hybridoma cells are isolated and separated into individual wells to assess the antigen-binding potential of the secreted antibodies.
The process of B-cell fusion is inefficient. Many B-cells are lost during this process, decreasing the probability of discovering a rare antibody that binds difficult-to-target epitopes. In contrast, our single B-cell screening method maximizes the B-cell isolation and characterization of antibodies secreted from B-cells increasing the probability of discovering rare antibodies that bind a target antigen with precision.

Question 43

What are the challenges of Single B cell Isolation?

Accepted Answer

Picking the right team of antibody experts to isolate single B-cells for monoclonal antibody production is critically important given the numerous challenges involved in B-cell isolation. First, the ability to make custom antigens, including conformation-dependent antigens, is critical to success. These antigens are not only necessary for the immunization steps but also for the isolation of antigen-specific B-cells by FACS. The synthesized antigen must be a specific purity and concentration for optimal success.
Successful B-cell isolation also requires an array of properly calibrated precision instruments to isolate B-cells and verify antigen binding capacity. In addition, the ability to culture naive and memory B-cells requires well-optimized conditions to promote cell proliferation and maintain cell viability. These parameters alone require years of optimization and expertise.
At ProteoGenix, our antibody experts have an average of 25 years of experience in antigen synthesis and antibody development. With 3 therapeutic antibodies on the market and over 30 antibodies in the preclinical and clinical phases of investigation, we know how to make high-performing antibodies for our clients. Book a call with one of our antibody experts to learn how we can help you accelerate your project needs.

Question 44

Which expression systems do you offer (CHO, HEK, E. coli, insect, yeast…)?

Accepted Answer

We offer a full panel of expression platforms to match your protein and application. This includes mammalian systems (CHO and HEK), with our proprietary XtenCHO® Race CHO line delivering top-tier performance for demanding projects. We also provide expression in insect cells, yeast, E. coli, and Bacillus subtilis for secreted proteins or specific production needs.

Question 45

How do I determine which system is best for my protein (size, glycosylation, solubility, function…)?

Accepted Answer

The optimal system depends on your protein’s characteristics and intended use. For example, mammalian systems are preferred when native folding or post-translational modifications are required, while E. coli can be the fastest and most cost-effective option for simpler proteins and short timelines. In all cases, our PhD-level experts will review your target and constraints with you and recommend the most suitable strategy.

Question 46

Can you express challenging proteins such as membrane proteins, secreted proteins, or multi-domain constructs?

Accepted Answer

Yes. We often handle complex targets, including membrane proteins, secreted proteins, and multi-domain constructs. Mammalian expression, especially with XtenCHO Race, is particularly well suited for difficult proteins. With over 1,500+ projects completed, our team has built deep expertise on hard-to-express targets and is recognized as a market reference for complex antibody and protein projects.

Question 47

Do you provide codon optimization or sequence engineering before expression?

Accepted Answer

Yes, we always optimize codons using our proprietary AI-powered tool to maximize expression efficiency. Additionally, we can analyze your sequence to identify any potential liabilities and can propose targeted mutations to improve developability and performance, ensuring your protein is ready for successful production.

Question 48

What scale options are available (small-scale screening, mid-scale, gram-scale)?

Accepted Answer

We support every scale, from high-throughput screening and small-scale expression to mid-scale production and gram-level manufacturing. Depending on your needs, we can run large-scale transient expression in bioreactors or develop stable cell lines for long-term, high-volume production.

Question 49

What purification techniques do you use (affinity, ion exchange, SEC, tag-based…)?

Accepted Answer

We use a range of purification techniques to ensure the highest quality protein. Our routine methods include Nickel (His-tag), Streptactin, antigen-affinity, Protein A/G, Ion Exchange (IEX), and Size Exclusion Chromatography (SEC). It can be tailored based on your protein’s specific needs.

Question 50

Do you offer additional services like tag removal, refolding, or buffer optimization?

Accepted Answer

Yes, we offer a variety of additional services to ensure optimal protein performance. These include tag removal, refolding, buffer optimization, endotoxin removal, stability testing, and concentration up to 50mg/mL, all tailored to meet your project requirements.

Question 51

What QC assays do you provide (SDS-PAGE, SEC-HPLC, endotoxin, concentration, activity tests…)?

Accepted Answer

We perform a comprehensive set of quality control assays, including SDS-PAGE, ELISA, FACS, SPR/BLI, SEC-HPLC, nanoDSF, MS, endotoxin assays, and concentration determination by UV280/Bradford, to ensure your protein meets the highest standards.

Question 52

What are your typical timelines for transient versus stable expression?

Accepted Answer

Transient expression projects can start from 15 business days, depending on the system and complexity. Stable cell line development typically requires around 4.5 months, reflecting the time needed to generate and validate high-performing stable clones.

Question 53

Can I supply my own vector, sequence, or cell line?

Accepted Answer

Yes, you can supply your own vector or sequence, and we can handle the vector construction for you. However, we use our own optimized cell lines for expression to ensure maximum efficiency and consistency.

Question 54

How flexible is your workflow? Can you adapt it to my preferred tags, buffers, and conditions?

Accepted Answer

Our services are highly customizable. We can adapt our workflow to suit your protein’s specific needs, including your preferred tags, buffers, and conditions, ensuring the best possible outcome for your project.

Question 55

Can you ensure endotoxin-free preparations for cell-based or in vivo studies?

Accepted Answer

Yes, we can guarantee endotoxin-free preparations with levels as low as <0.1 EU/mL.

Question 56

What documentation accompanies each batch?

Accepted Answer

Each batch is accompanied by a Certificate of Analysis (CoA), a production report with quality control data, and a packing slip, providing full traceability and transparency for your project.

Question 57

Do you provide scalable solutions from discovery-grade to preclinical-grade protein production?

Accepted Answer

Yes. As part of our therapeutic antibody discovery services, we can support early discovery programs with high-throughput antibody production service to screen and select candidates, then scale up into larger-volume production under strict process control with custom cell line development. For preclinical applications, we also provide enhanced purity, QC, and documentation to meet your functional and regulatory expectations.

Question 58

How do you guarantee purity, stability, and activity of the final protein?

Accepted Answer

While we guarantee yield and purity based on SDS-PAGE and SEC-HPLC after a pilot run, activity for antibodies is also guaranteed based on ELISA results. We ensure the highest quality proteins, optimized for your application, after performing thorough testing and validation.

Question 59

Can I request pilot runs before committing to large-scale production?

Accepted Answer

Yes, we always conduct pilot runs to determine the optimal conditions, yield, and purity before committing to large-scale production.

Question 60

Can you support downstream applications such as assay development, antibody generation, or structural studies?

Accepted Answer

Yes, we can support assay development such as ELISA and antibody generation, leveraging our expertise in hybridoma, phage display, and single B cell technologies. However, we do not conduct structural studies at this time.

Question 61

What post-project support do you offer (storage, re-expression, additional purification steps…)?

Accepted Answer

We offer a wide range of post-project support, including storage, production optimization, scale-ups, purification polishing, sequence optimization (e.g., affinity maturation and developability improvement), and stable cell line development, ensuring your protein can be used long-term and in future applications.

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