Donkey Antibody Library Construction by Phage Display

Introduction

Phage display technology is a powerful tool for generating diverse antibody libraries, including those derived from non-human species. Among the various animal models, donkeys have recently emerged as a valuable source for antibody discovery due to their unique immunological characteristics. At Creative Biolabs, we offer a comprehensive donkey antibody library construction by phage display service, utilizing advanced phage display systems to generate high-quality, diverse antibody libraries tailored for your research.

Donkey as an Immunization Host

Donkeys (Equus asinus) offer unique immunological benefits for monoclonal antibody development, often outperforming mice or rabbits.

• Strong immunity: As large mammals, they mount robust, diverse responses, recognizing diverse epitopes—including those on complex antigens (e.g., multi-subunit proteins, glycoproteins, post-translationally modified structures)—ideal for challenging targets.
• High serum yield: They produce large volumes of high-titer serum, supplying ample immunoglobulin for downstream use, reducing variability in screening/validation.
• Broad epitope recognition: Their immune systems detect a wide range of epitopes, even conserved ones, yielding diverse antibody repertoires to uncover novel antibodies missed in other species.

Antibody Types from Donkey Libraries

The donkey immune system provides access to various antibody types that are beneficial for research. These include IgG, IgM and IgA antibodies. The most common class of antibody used in therapeutic applications, donkey-derived IgGs can be isolated and purified from serum. These antibodies are typically isolated after immunization and can be further engineered into various formats, such as Fab and scFv, for specific applications. IgM antibodies, which are the first antibodies produced in response to an infection or foreign antigen, can also be harvested from donkey serum. These antibodies are often used in research on immune responses or for developing diagnostic tests due to their ability to recognize antigens with high avidity. IgA antibodies, which play a critical role in mucosal immunity, can be isolated from donkey mucosal samples. These antibodies are particularly useful for targeting antigens that are located at mucosal surfaces, such as those found in respiratory or gastrointestinal diseases.

Services for Donkey Antibody Library Construction by Phage Display

At Creative Biolabs, we provide a comprehensive set of customizable services for donkey antibody library construction, designed to meet your specific research and therapeutic needs. We work with various species-specific antibody formats, including scFv, Fab, and IgG, and offer flexible solutions based on immunization, naïve, semi-synthetic, and synthetic libraries. Our cutting-edge phage display technology ensures the highest diversity and specificity, enabling efficient discovery of monoclonal antibodies against challenging targets.

Phage Display Technology: An Overview

Phage display is a technique that involves the presentation of peptides, proteins, or antibodies on the surface of bacteriophages. The peptides or proteins are genetically linked to one of the phage coat proteins, allowing for the identification of specific binders through a process called biopanning. By screening vast libraries of phage-displayed proteins, researchers can identify high-affinity antibodies that bind specifically to a target of interest. There are three commonly phage types used in display:

• M13 phage is widely used for antibody display due to its stable, filamentous structure, and it allows the display of up to 2,700 copies of the antibody on the phage surface
• T4 and T7 phages can provide additional versatility with the ability to display larger proteins and increased resistance to harsh screening conditions.

Phage display libraries are incredibly useful because they can hold up to 10¹¹ unique peptides or antibodies—that's a huge range, which makes thorough screening much easier. They're also really flexible: you can choose from different types like immune, naïve, semi-synthetic, or synthetic antibody libraries, so they fit all kinds of research needs. On top of that, their diversity—take donkey antibody libraries, for example—makes them powerful tools. They even help find antibodies for new antigens, including those that don't trigger a strong immune response.

Donkey Antibody Library Construction Process

Step 1: Antibody Gene Collection

The first step in constructing a donkey antibody library is the collection of the antibody genes. The genes encoding donkey antibodies are isolated from the peripheral blood lymphocytes of donkeys. These lymphocytes are carefully extracted, and the RNA is subsequently isolated. The antibody repertoire is represented by heavy and light chain variable regions that are amplified to generate a diverse pool of antibody genes.

Step 2: Cloning the Antibody Genes into Phagemid Vectors

Once the antibody genes are obtained, they are cloned into phagemid vectors. These vectors are modified versions of bacteriophages that allow the display of antibody fragments (e.g., scFv or Fab) on the surface of the phage. The genes encoding the donkey antibodies are fused with the phage's coat protein, enabling the creation of a large, diverse library of phage particles displaying unique donkey antibodies.

Step 3: Library Amplification

The cloned phage library is then amplified inE. coli bacteria, producing high concentrations of the phage particles that display the donkey antibodies. This amplification step ensures that a large number of phage particles are available for the screening process.

Step 4: Screening and Biopanning

Biopanning is the key step in the phage display process, where the antibody library is screened to identify phage particles that bind specifically to a target antigen. During biopanning, the phage library is incubated with an antigen of interest, and non-binding phages are washed away. The remaining phages that display antibodies with high affinity for the target are then eluted, amplified, and subjected to further rounds of selection to increase the binding specificity.

Step 5: Antibody Characterization and Sequencing

After the screening process, the selected high-affinity phages are analyzed and sequenced to identify the unique antibody sequences. These antibodies can then be further characterized for their affinity, specificity, and potential for use in therapeutic or diagnostic applications.

Fig.1 Five phage display selection steps.¹

Why Choose Creative Biolabs for Donkey Antibody Library Construction?

At Creative Biolabs, we leverage years of expertise in phage display technology to deliver top-quality donkey antibody libraries for a wide range of applications. Our service includes:

• We provide custom antibody libraries with high diversity and specificity, ensuring the best possible chance of finding high-affinity binders.
• Our tailored biopanning strategies, including solid-phase, in-solution, and cell-based screening, ensure that we can efficiently isolate the best binders for any given target.
• Our high-throughput screening capabilities allow for fast and efficient identification of the most promising antibody candidates.

Applications of Donkey Antibody Libraries

Donkey-derived antibodies offer several unique advantages for research purposes:

• Donkey antibodies can be used to generate monoclonal antibodies for treatment of diseases such as cancer and autoimmune disorders.
• Donkey antibodies are suitable for developing diagnostic assays to detect specific biomarkers in various diseases.
• Donkey antibodies can be utilized to explore biological pathways and identify new therapeutic targets.

Creative Biolabs' donkey antibody library construction by phage display service provides a powerful tool for the discovery of unique and high-affinity antibodies. By utilizing state-of-the-art phage display technology and our extensive expertise, we are able to offer a comprehensive and efficient solution for antibody development. If you are looking for antibodies for research, our service ensures you receive the best quality and most reliable results.

Reference:

1. Ledsgaard, Line, et al. "Basics of antibody phage display technology." Toxins 10.6 (2018): 236. Distributed under Open Access license CC BY 4.0, without modification. https://doi.org/10.3390/toxins10060236

Please kindly note that our services can only be used to support research purposes (Not for clinical use).