Monoclonal antibody (mAb) discovery
Monoclonal antibodies (mAbs) originate from one unique parent cell, thus binding only to a single epitope. Monoclonal antibody discovery typically refers to the screening and identification of specific antibodies that target a specific epitope for the diagnosis and treatment of diseases.
There are several approaches to generating therapeutic mAbs. Two of the most common production methods are hybridomas and phage display.
mAb production via hybridomas
In this approach, scientists fuse immortalized myeloma cells with splenocytes obtained from animals previously exposed to the antigen of interest. The resulting fused cell is called a hybridoma. The advantage of a hybridoma is that it now possesses the ability to produce mAbs while dividing indefinitely, whereas splenocytes have a finite lifespan. Researchers then screen thousands of hybridomas to identify the optimal ones based on antigen specificity and immunoglobulin class. These candidates are subsequently analyzed, characterized, and scaled up for downstream processes.
mAb production via phage display
In this method, the variable heavy and light genes of an antibody are introduced into a bacteriophage coat protein gene, resulting in phages that display the antibody on their surface. Using this strategy, scientists can generate an antibody library displaying millions of different antibodies. These displaying phages are then screened against the antigen of interest, and those that are highly specific are identified, isolated, and further characterized for downstream work.
How are monoclonal antibodies produced?
The traditional monoclonal antibody (mAb) production process usually starts with generation of mAb-producing cells (i.e. hybridomas) by fusing myeloma cells with desired antibody-producing splenocytes (e.g. B cells). These B cells are typically sourced from animals, usually mice. After cell fusion, large numbers of clones are screened and selected on the basis of antigen specificity and immunoglobulin class. Once candidate hybridoma cell lines are identified, each "hit" is confirmed, validated, and characterized using a variety of downstream functional assays. Upon completion, the clones are scaled-up where additional downstream bioprocesses occur.
Additional resources for antibody discovery
Accelerate your monoclonal antibody discovery with our product portfolio specifically designed to shorten cell line development time and deliver stable clones with higher affinities and expression levels.
-
Cell line Development
Cell line development is a critical step in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. The process often begins with transfecting the host cell type with the DNA encoding the therapeutic protein of interest allowing for random or directed integration of target DNA into the host cell genome. Thousands of clones are screened to isolate the rare high producing cells, a manual and time-consuming process.
Cell line Development
Cell line development is a critical step in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. The process often begins with transfecting the host cell type with the DNA encoding the therapeutic protein of interest allowing for random or directed integration of target DNA into the host cell genome. Thousands of clones are screened to isolate the rare high producing cells, a manual and time-consuming process.
Comparison of traditional cloning methods vs. f.sight
In this study, we conducted six sets of experiments, cloning two CHO cell lines with animal-component-free (ACF) cell culture media and supplements, and compared the performance a CloneSelect™ f.sight single cell printer versus two other accepted cloning methods (FC and LD).
View our introductory video and download application note.
Comparison of traditional cloning methods vs. f.sight
In this study, we conducted six sets of experiments, cloning two CHO cell lines with animal-component-free (ACF) cell culture media and supplements, and compared the performance a CloneSelect™ f.sight single cell printer versus two other accepted cloning methods (FC and LD).
View our introductory video and download application note.
-
Confident identification of monoclonal CHO-S cells
Growing CHO cells in CloneMedia CHO Growth A is a more efficient way to clone CHO cells than performing limiting dilution in liquid media. Semi-solid media immobilizes the cells which prevents the cells from moving during routine handling. This enables researchers to confidently track the growth of a single cell into a colony.
Confident identification of monoclonal CHO-S cells
Growing CHO cells in CloneMedia CHO Growth A is a more efficient way to clone CHO cells than performing limiting dilution in liquid media. Semi-solid media immobilizes the cells which prevents the cells from moving during routine handling. This enables researchers to confidently track the growth of a single cell into a colony.
eBook: Optimize your high value cell lines
In this eBook, we present an overview of the cell line development workflow as well as high throughput solutions for accelerating the process, and enabling easier and faster selection of high-producing mammalian cell lines.
eBook: Optimize your high value cell lines
In this eBook, we present an overview of the cell line development workflow as well as high throughput solutions for accelerating the process, and enabling easier and faster selection of high-producing mammalian cell lines.
-
Enhanced development of virus-specific hybridomas
Double-stranded DNA (ds-DNA) viruses cause a variety of disease in humans. Species within the order of Herpesvirus and the families Adenovirus and Papillomavirus can cause symptoms in humans with similar presentations. Furthermore, serological and protein-based diagnostic confirmation for a particular virus may be complicated due to antigen conservation within viral genomes. A high degree of proteomic homology and serum cross reactivities lead to poor differentiation between individual species of ds-DNA viruses. The purpose of the research was to identify an optimal producer cell line that secrets a highly specific, non-cross-reactive monoclonal antibody to be used in biotherapeutics development.
Enhanced development of virus-specific hybridomas
Double-stranded DNA (ds-DNA) viruses cause a variety of disease in humans. Species within the order of Herpesvirus and the families Adenovirus and Papillomavirus can cause symptoms in humans with similar presentations. Furthermore, serological and protein-based diagnostic confirmation for a particular virus may be complicated due to antigen conservation within viral genomes. A high degree of proteomic homology and serum cross reactivities lead to poor differentiation between individual species of ds-DNA viruses. The purpose of the research was to identify an optimal producer cell line that secrets a highly specific, non-cross-reactive monoclonal antibody to be used in biotherapeutics development.
Hybridoma selection using HAT medium
Hybridoma selection after fusion of myelomas and spleen cells is a critical step in monoclonal antibody production. Often scientists use the HAT (hypoxanthine-aminopterin-thymidine) method to accomplish this task.
Hybridoma selection using HAT medium
Hybridoma selection after fusion of myelomas and spleen cells is a critical step in monoclonal antibody production. Often scientists use the HAT (hypoxanthine-aminopterin-thymidine) method to accomplish this task.
-
Hybridoma technology
Hybridoma technology is a method for mass-producing antibodies in a hybrid cell line generated from the fusion of antibody-producing B-cells with an immortalized myeloma cell line, now called a hybridoma cell. Because every B-cell produces a unique antibody, single-cell cloning of hybridomas can be used to generate a diverse library of unique monoclonal antibodies at a large scale, which are very frequently used in the prevention, diagnosis, and treatment of disease.
Hybridoma technology
Hybridoma technology is a method for mass-producing antibodies in a hybrid cell line generated from the fusion of antibody-producing B-cells with an immortalized myeloma cell line, now called a hybridoma cell. Because every B-cell produces a unique antibody, single-cell cloning of hybridomas can be used to generate a diverse library of unique monoclonal antibodies at a large scale, which are very frequently used in the prevention, diagnosis, and treatment of disease.
Monoclonality assurance
Cell line development and assurance of monoclonality are critical steps in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. A cell line can be established following the isolation of a single viable cell robustly expressing the protein of interest. A key milestone in this process is documenting evidence of clonality. Documentation of clonality is typically image-based, whereby an image of a single cell is produced and included in regulatory filings.
Monoclonality assurance
Cell line development and assurance of monoclonality are critical steps in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. A cell line can be established following the isolation of a single viable cell robustly expressing the protein of interest. A key milestone in this process is documenting evidence of clonality. Documentation of clonality is typically image-based, whereby an image of a single cell is produced and included in regulatory filings.
-
Phage display
Phage display is a technique used to study the interaction of proteins displayed on the surface of a bacteriophage with other molecules such as peptides, DNA, and other proteins. Phage display is commonly used to find high-affinity interactions between antibodies and antigens, which play a critical role in viral pathogenesis, vaccines, and other treatments.
Phage display
Phage display is a technique used to study the interaction of proteins displayed on the surface of a bacteriophage with other molecules such as peptides, DNA, and other proteins. Phage display is commonly used to find high-affinity interactions between antibodies and antigens, which play a critical role in viral pathogenesis, vaccines, and other treatments.
Preparing and plating cells in semi-solid medium
Plating cells in the viscous semi-solid medium may be challenging to first-time users. Watch our 4-minute tutorial to learn some tips and tricks on how to plate cells in this medium more efficiently.
Preparing and plating cells in semi-solid medium
Plating cells in the viscous semi-solid medium may be challenging to first-time users. Watch our 4-minute tutorial to learn some tips and tricks on how to plate cells in this medium more efficiently.
Resources of Monoclonal Antibodies (mAbs)
eBook
Optimize your high-value cell lines
Optimize your high-value cell lines
During the last decade, production of monoclonal antibodies (mAbs) and recombinant proteins using mammalian cells has led to a boom in the introduction of biotherapeutic proteins to the m…
Application Note
Comparison of traditional cloning methods vs. CloneSelect Single-Cell Printer f.sight using CHO cell lines commonly used for monoclonal antibody production
Comparison of traditional cloning methods vs. CloneSelect Single-Cell Printer f.sight using CHO cell lines commonly used for monoclonal antibody production
As regulations for cell line development become increasingly more stringent, researchers will be required to perform single-cell cloning and provide evidence that a cell line is derived…
-
Application Note
Confident Identification of Monoclonal CHO-S Cells Grown in Semi-Solid Media using the CloneSelect Imager
Confident Identification of Monoclonal CHO-S Cells Grown in Semi-Solid Media using the CloneSelect Imager
Regulatory criteria to obtain approval of biological drugs are constantly evolving. One of the regulatory metrics for biological cell lines, the digital documentation of monoclonality, has…
-
Application Note
Enhanced development of virus-specific hybridomas using ClonePix and CloneSelect Imager Technologies
Enhanced development of virus-specific hybridomas using ClonePix and CloneSelect Imager Technologies
The ClonePix System was employed in the high-throughput screening of hundreds of sub-cloned colonies from parental hybridoma material (historically parental line yields were less than 1 mg/…
Videos & Webinars
Hybridoma selection using HAT medium
Preparing and Plating Cells in Semi-Solid Medium