There have been remarkable advancements in antibody generation technology over the years, offering scientists many techniques to generate antibodies for various applications ranging from discovery research to diagnostics and therapeutics. Three prominent methods for generating monoclonal antibodies are phage display, hybridoma, and single plasma B cell cloning.
Let’s explore the differences between these methods and provide insights into how to choose the right one for your specific project.
Phage display is a powerful technique for in vitro antibody selection. Bacteriophages (viruses that infect bacteria) are used to express antibody fragments, typically single-chain variable fragments (scFv) or Fab fragments, on their surface. These displayed antibody fragments can then be screened for binding to a specific target of interest.
Diversity: Phage display libraries can contain a vast number of different antibody clones, allowing for a wide range of potential binders to be screened. Additionally, toxic antigens can show up during screening and be potentially screened, which is more difficult in a human or animal.
Speed: The selection process in phage display can be relatively quick, making it suitable for rapid antibody development.
Limited to antibody fragments: Phage display typically produces antibody fragments rather than full antibodies, which may not be suitable for all applications.
In vitro process: It does not produce antibodies in their native, post-translational modified form, which may lead to developmental issues, aggregation, and instability.
Lower affinity hits: A naïve library may lead to hits with lower affinities compared to an immunized library source.
When to choose Phage Display
- If you need to identify antibody fragments more quickly than hybridoma would allow for.
- When working with challenging targets that are hard to immunize against.
Working with a CRO for Phage Display
A CRO specializing in phage display can provide valuable expertise and resources for library construction, screening, and optimization. They can accelerate the process, ensuring you obtain high-quality antibody fragments tailored to your project.
Hybridoma technology is a classic method for generating monoclonal antibodies. The fusion of a specific B cell (producing the desired antibody) with a myeloma cell creates a hybrid cell line that continuously produces the antibody of interest.
Full antibodies: Hybridoma technology produces intact monoclonal antibodies with all the native properties.
High specificity and affinity: Antibodies generated typically exhibit high binding specificity and affinity.
Time-consuming: The process of generating hybridomas and screening them for desired antibodies can be time-consuming, especially if animals other than mice are being used.
Non-mice model difficulties: If you use animals besides mice, creating a hybridoma is very difficult due to lack of fusion partner cells (melanoma in mice).
Limited diversity: The immune system’s repertoire may not include antibodies against all antigens.
When to choose Hybridoma Technology
- When you require full antibodies with native properties.
- If you have access to the target antigen or a closely related antigen for immunization.
Working with a CRO for Hybridoma Technology
A CRO experienced in hybridoma can assist in antigen preparation, immunization strategies, and screening processes. Their expertise can streamline antibody production and help you obtain highly specific monoclonal antibodies efficiently.
Single B Cell Cloning
Single Plasma B cell cloning is a relatively recent advancement in antibody production. Isolation and cloning of individual B cells produce antibodies against a specific target. This method allows for the generation of monoclonal antibodies with minimal manipulation and the retention of the natural antibody repertoire.
Speed: Specificity to targets and some functionality can be screened on the same day they are harvested from the immunized animal.
Native antibodies: Single B cell cloning produces antibodies in their native form, maintaining their natural post-translational modifications.
High specificity: The antibodies generated are often highly specific to the target antigen.
Technically demanding: Isolating and cloning single B cells can be technically challenging and may require specialized equipment and expertise such as the Beacon Optofluidic System®.
Requires robust assays: For functional screening, robust assays are required. These can sometimes be demanding to develop or require a highly trained scientific team.
When to choose Single B Cell Cloning
- When you require native antibodies with high specificity and minimal manipulation.
- If working with a limited sample source or rare B cells.
- When cross-reactivity or functional activity (blocking ligand, etc) are important to know early on.
Working with a CRO for Single B Cell Cloning
Success with b cell cloning requires an experienced and thorough team. CROs specializing in single plasma B cell cloning can efficiently handle the isolation, cloning, and validation processes. This increasing the likelihood of successful antibody generation, especially when dealing with limited or challenging sample sources. We understand that precision and accuracy are paramount to delivering top-notch results to our clients.
Choosing the Right Method for Your Project
The choice between phage display, hybridoma technology, and single B cell cloning depends on several factors, including:
- Project Goals: Consider whether you need full antibodies or antibody fragments and the level of specificity and affinity required.
- Target Antigen: Assess the nature of your target antigen and its availability for immunization or screening.
- Resources: Evaluate the availability of equipment, expertise, and funding for the chosen method.
- Timeline: Consider the urgency of your project, as some methods may be quicker than others.
Each antibody production method has its strengths and limitations, and the choice should align with the specific needs and constraints of your project. Working with a Contract Research Organization (CRO) that specializes in your chosen method can significantly enhance your antibody production process by providing expertise, resources, and streamlined workflows. This collaborative approach ensures that you obtain high-quality antibodies tailored to your research or application, ultimately accelerating your scientific endeavors.
If you’re still not sure which route is appropriate for your project, contact our team to discuss your specific project with a scientist.