An Introduction to Absolute Antibody Custom Services

Absolute Antibody has been a trusted name in recombinant antibody technology for over a decade. Our catalog contains more than 13,500 off-the-shelf recombinantly produced antibodies that guarantee continued functionality and batch-to-batch reproducibility. The recombinant technologies we use to manufacture our antibody catalog are also available to clients worldwide as custom antibody services. From sequencing to engineering and expression, Absolute Antibody will be your consultative partner for all your antibody needs.

Antibody Sequencing

Knowing your hybridoma or antibody protein sequence is crucial for the security and continuation of your antibody production. Antibody-producing hybridomas are susceptible to many pitfalls, including:

• Cell line instability
• Diminished antibody production
• Genetic drift and isotype switching
• Non-monoclonality (Bradbury et al, 2018)
• Cross contamination and cell line death

Absolute Antibody’s sequencing services help mitigate the risks to your hybridomas and antibodies and provide a back-up plan should catastrophe strike.

Hybridoma Sequencing

• Novel NGS approach to high-throughput sequencing
• Compatible with any species and isotype
• Identifies all heavy chain and light chain sequences and quantifies their relative abundance
• Rescue contaminated or dying hybridomas

Antibody Protein Sequencing

• Sequence from purified monoclonal antibody when hybridoma is not available
• Combines mass spectrometry with high accuracy de novo peptide sequencing and recombinant expression

Sequencing can be performed as a stand-alone activity, or as the first step toward antibody engineering and transient serum-free expression. Whether it is a hybridoma or purified monoclonal antibody protein, Absolute Antibody offers industry-leading sequencing services to help safeguard your invaluable antibody. We have successfully sequenced more than 5,000 hybridomas and counting.

Antibody Engineering

Antibody engineering is the process of reformatting an antibody in a variety of different ways. Engineering services offered by Absolute Antibody include:

• Species switching- changing the antibody backbone to that of a different animal
• Class switching- altering the isotype or subtype of an antibody
• Reformatting- creating antibody fragments, multispecific antibodies, and changing the Fc region

Antibody Reformatting

Antibody Fragments

Antibody fragments are often utilized when Fc-mediated effects are not required and a smaller molecule is beneficial, such as anti-cancer antibodies for tumor penetration.

Bispecific Antibodies

Bispecific and multispecific antibodies are growing in popularity, especially in therapeutics. These antibodies combine multiple epitopes into a single antibody structure and offer several benefits:

• Effector cell recruitment to target molecules
• Drug delivery to a specified target
• Dual-target antibodies for increased cell type selectivity

Fc Domain Engineering

Fc domain engineering can be useful for multiple reasons. Absolute Antibody can alter the Fc region to mediate effector function and extend or shorten the plasma half-life. In addition, Fc regions can be silenced by abolishing binding to FcγR using our Fc Silent™ mutation or mAbsolve’s STR mutations.

The changes that can be made to an antibody may seem overwhelming, but our team of experts will work with you to determine the best course of action to reach your desired outcome. For a more in-depth look at antibody engineering, including information about antibody humanization, download our Guide to Recombinant Antibody Engineering white paper.

Antibody Expression

Antibodies can be expressed using different methods, depending on the type of antibody being produced.

• Polyclonal antibodies are made from the immunization of animals.
• Monoclonal antibodies were traditionally produced using hybridomas.
• Recombinant monoclonal antibodies can be produced transiently, in a pool, or in a stable cell line.

Absolute Antibody’s expression services transiently produce recombinant antibodies in serum-free mammalian HEK and CHO cell lines. This animal-free production method ensures:

• Milligram-to-gram scale of antibody production
• Batch-to-batch consistency and reproducibility
• High purity and low endotoxin levels

Absolute Antibody has experience manufacturing antibodies for pharmaceutical and biotechnology companies, diagnostic assay developers, and academic researchers. All production occurs at our ISO 9001:2015 certified manufacturing facility located in the UK. We have produced more than 21,000 different recombinant antibodies in more than 180 engineered formats. Let us be your consultative partner and work together for a successful custom antibody project.

For more information about our custom services, please contact us.

Additional Resources

Guide to Recombinant Antibody Engineering – This white paper provides an overview of recombinant antibody engineering, covering key antibody background and describing different design parameters to consider when engineering an antibody.

Periodic Table of Antibodies – This poster showcases 180 different antibody formats Absolute Antibody has engineered and produced for our clients.

Why Go Recombinant? – In this presentation, Absolute Antibody CSO Michael Fiebig, PhD, discusses the significant benefits of recombinant antibodies compared to traditional hybridoma-produced monoclonal antibodies.