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^-12m) 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.

DATA REPORT

Get free access to genuine, representative data
from a Rabbit Single B-Cell Screening project

An ideal example of the insights you can expect from this type of report

This comprehensive report includes:

  • A full set of detailed data
  • Advanced tests and results
  • Key insights at every step of the process

Download the Data Report – protein antigen

Download the Data Report – peptide antigen

Why Choose ProteoGenix’s Single B Cell Sequencing Service?

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.
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ProteoGenix’s Single B cell sequencing services FAQ

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.

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.

 

The Evolution of Monoclonal Antibody Development

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.

More antibody Discovery and Expression services

Hybridoma development Services – ProteoGenix
Recombinant Antibody Services – ProteoGenix
Phage Display Custom Services – Proteogenix

 

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