Single B Cell Sequencing Service – ProteoGenix

Unlock the full potential of native-like antibodies with ProteoGenix’s Custom Single B Cell Sequencing Service. With the RocketAb immunization protocol, revolutionize your mAb development by offering a fast turnaround—saving up to 4 weeks compared to traditional methods and using 50% less antigen. Choose from mouse and rabbit species to best suit your project needs, all while ensuring maximum B cell diversity and the preservation of natural VL/VH pairing. Our innovative adjuvant promotes native protein recognition without the need for emulsification, setting a new benchmark for efficiency and specificity in single B cell cloning services. Guaranteeing at least three unique, high-affinity (10^-12 M) clones, ProteoGenix accelerates your path to breakthrough discoveries with high-quality, high-affinity monoclonal antibodies. Opt for ProteoGenix and transform your antibody development challenges into pioneering solutions with our leading-edge custom B cell sorting services.

Why choose ProteoGenix for your single B-cell sorting?

Fast Turnaround

Save up to 4 weeks with our RocketAb protocol.

Less Antigen Required

Use 50% less antigen for immunizations.

Choice of Species

Mouse and rabbit options for mAb development.

Native Antigen Recognition

Our adjuvant ensures no emulsification needed.

High-Affinity Clones Guaranteed

Receive at least 3 clones with 10^-12 M affinity

Advanced B Cell Sorting

Utilizing FACS for precise antigen-specific isolation.

Proprietary High-Performance Cells

For efficient antibody gene expression.

IP-Free Guarantee

Full ownership of your research outcomes.

High Throughput Screening

Expedite discovery with our advanced platforms.

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Our Single B Cell Sequencing Service Process

Immunization: 4-6 weeks

Antigen validation with SDS-PAGE: using either client-provided or ProteoGenix-designed antigens.
Rabbit& mouse immunization leveraging the RocketAb protocol: for optimized antigen response saving up to 4 weeks

Single B-cell Sorting and Screening: 2-3 weeks

PBMCs and spleen lymphocytes isolation, followed by precise antigen-specific single B-cell sorting via FACS (1 cell/well).
Cultivation of B-cells and antigen-specific screening using ELISA or client’s specific application like WB, IP or IHC

Recombinant Mouse & Rabbit Antibody Production and Screening: 2 weeks

Sequencing of the highest affinity clones.
Transient expression in our high-performance XtenCHO™ system.
Final ELISA screening of supernatant for antigen binding

Custom Solutions and Additional Services

Custom Antigen Design and Synthesis: Offer details on custom solutions, including antigen design, synthesis, isolation, and purification to meet specific project needs.
Crossreactivity and Neutralization Assessments: Introduce additional services like assessing the crossreactivity profile of specific antibodies or identifying clones with neutralization capabilities.

Delivrables

Supernatant Samples: Up to 30 clones post-screening; up to 10 recombinant clones post-production, or purified antibody samples in PBS pH7.5.
Amino Acid Sequences: Variable regions of the top 3 positive clones, selected by the customer after sample testing and data comparison.

ProteoGenix’s Single B cell sequencing services FAQ

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.
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.
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.
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.
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.

The Science Behind Single B Cell Sequencing

Understanding B Cells and Their Role in Immunity

B Cell Biology: B cells, a cornerstone of the adaptive immune system, are specialized cells that produce antibodies to neutralize pathogens. Their unique ability to recognize foreign antigens makes them pivotal in the body’s defense mechanism. Through our custom single B cell screening service, we harness this natural defense to develop targeted antibodies for research and therapeutic use.
Antibody Diversity: The remarkable diversity of antibodies, essential for the immune system’s ability to combat a wide range of pathogens, is generated by the rearrangement of gene segments in B cells. This process results in antibodies with unique antigen specificities. Our single B cell sequencing service leverages this natural diversity, employing cutting-edge single B cell sorting and cloning techniques to isolate and clone antibodies with the desired specificities for your custom research needs.

The Evolution of Monoclonal Antibody Development

Traditional Methods vs. Single B Cell Sequencing

Traditional monoclonal antibody development methods, such as hybridoma technology, have been instrumental in advancing biomedicine, offering a solid foundation for producing a wide array of therapeutic antibodies. These methods have enabled significant discoveries and developments in the field. Complementing this, ProteoGenix’s single B cell sequencing service introduces a contemporary approach that further enhances the precision and efficiency of antibody discovery. Below is a table comparing the three methods used for monoclonal antibody development.

What are the main differences between single B-cell sorting and other monoclonal antibody screening methods?

To date, several techniques have been developed to isolate and screen monoclonal antibodies (mAbs). This table summarizes the main advantages and drawbacks of each technique.

	Pro	Cons
Hybridoma	High antibody affinity Preservation of natural VH-VL pairing (positively impacts developability) Cheap	High loss of diversity due to low efficiency of cell fusion Limited to rodents as myeloma cells not efficient in other species Humanization needed Not adapted to antigens with low immunogenicity and/or toxicity Animal use
Phage Display	No need for humanization thanks to fully human libraries & humanized mice Maximized diversity due to random VH-VL pairing High-throughput screening allowing identification of multiple binders Adapted to antigens with low immunogenicity and/or toxicity Very fast No animal use thanks to premade libraries	Non-natural VH-VL pairing can negatively impact developability Potential bias due to phage particle Low/medium antibody affinity due to lack of immunizations (naïve libraries) High affinity antibodies expensive & long to develop as require custom immune library construction
Single B-Cell Sorting	High antibody affinity Preservation of natural VH-VL pairing High-throughput screening Best for discovery of rare antibodies against challenging targets (e.g. conformational epitopes) with maximized developability	Not adapted to antigens with low immunogenicity and/or toxicity

Advantages of Single B Cell Sequencing

One of the key benefits of single B cell sequencing is its ability to preserve the natural pairing of heavy and light chains of antibodies. This preservation is crucial for maintaining the native function and specificity of antibodies, which is often compromised in traditional methods. Our custom single B cell screening and single B cell cloning services are designed to leverage this advantage, providing researchers with high-fidelity antibodies for their projects.

B cell sorting and its significance in various fields

Clinical applications: B cell sorting is pivotal in identifying potent antibodies for therapeutic applications, speeding up the development of treatments for a wide range of diseases. Our single B cell sorting service plays a crucial role in advancing personalized medicine.
Diagnostics: In diagnostics, the precision of single B cell sequencing enhances the development of highly specific diagnostic tools. This technology allows for the early detection of diseases, improving patient outcomes.
Research: For researchers, single B cell cloning and sequencing services open new avenues for understanding immune responses and discovering novel biomarkers. This technology is instrumental in exploring the vast diversity of the immune repertoire, facilitating groundbreaking discoveries in immunology.

Cutting-Edge Technologies in Single B Cell Sequencing

High-Throughput Sequencing: This technology enables the rapid and comprehensive analysis of B cell repertoires by sequencing millions of DNA molecules simultaneously. It’s crucial for identifying a wide array of antibody sequences within a sample, providing insights into the immune response and facilitating the discovery of novel antibodies with therapeutic potential.
Fluorescence-Activated Cell Sorting (FACS): FACS is a powerful tool for isolating individual antigen-specific B cells. By labeling cells with fluorescent markers and passing them through a laser, cells are sorted based on their fluorescence intensity. This method is instrumental in ensuring the high specificity of the antibodies produced, as it allows for the selection of B cells that are precisely targeted against specific antigens.

From Isolation to Antibody Production: A Closer Look

Isolation of Antigen-Specific B Cells: This process involves isolating B cells that produce antibodies against specific antigens. Techniques like fluorescence-activated cell sorting (FACS) are employed to select these cells with high precision, ensuring that only B cells with the desired specificity are chosen for further analysis.
Cloning and Expression: After isolation, the heavy and light chain genes from the selected B cells are cloned and expressed. This step is crucial for producing monoclonal antibodies. Proprietary cell lines, known for their high expression efficiency, are often used to ensure that the resulting antibodies are produced in sufficient quantities and retain their native functionality.