Custom antibody development FAQs

Some of the most common questions around our custom antibody generation services.

Overview


What services do you provide?

We provide comprehensive solutions for custom antibody development, from initial target evaluation all the way through to delivery of purified fit-for-purpose antibody for use in research, diagnostic or therapeutic programs.

We have highly experienced teams dedicated to antibody discovery for custom projects. We design and execute tailor-made discovery and selection strategies to generate and select binders that meet your desired antibody specifications.

What platforms do you use for custom antibody discovery?

We have three core antibody discovery platforms:

  1. RabMAb® rabbit monoclonal recombinant antibody development via either hybridoma or peripheral blood B-cell cloning techniques
  2. Next Generation Sequencing (NGS)-based rabbit monoclonal discovery
  3. AxioMx™ phage display using a human framework, initially with scFv but fully enabled for rapid IgG conversion.

The RabMAb and NGS platforms are based on rabbit immunization, and AxioMx employs an in vitro system. Selection of the most appropriate platform is dependent on the specific target, antibody requirements, and customer timelines. In some cases, multiple methodologies are used to maximize success.

Why rabbit?

Unique features of the rabbit immune system make it ideal for antibody generation, particularly to challenging targets.  Rabbits generate an expansive B-cell repertoire due to reliance on a gene-conversion mechanism in concert with more common somatic hypermutation during B-cell development. 

The rabbit’s immune system and varied antigen-presentation support diverse novel epitope recognition with less immunodominance than typically observed in other rodent hosts. This leads to the generation of a wide selection of antibodies with different binding properties and allows for differentiation between subtle structural variants such as post-translational modifications, point mutations, and epigenetic targets. The diverse epitope recognition found in the rabbit also helps in finding high-sensitivity antibody pairs.

An extra bonus is that rabbits have just one subclass of IgG, so there is no need for isotyping and antibodies can be compared directly at an early stage in screening cascades.

Can you give more information on your different platforms?

RabMAb antibody discovery

Hybridoma-based

Our RabMAb antibody discovery platform is built on rabbit hybridoma technology to generate highly specific, recombinantly-produced antibodies. We use our proprietary rabbit fusion partner bringing increased stability compared to hybridoma generated with heterogenous myeloma.

Selection is applied to the crude B-cell population from the animal to enrich for target specific B-cells before the hybridoma fusion step and maximize the number of target-specific clones for evaluation. This B-cell enrichment, in combination with the broad epitope diversity yielded by the rabbit immune response, maximizes the testable epitope space, increasing the opportunity of discovering antibodies that meet challenging specificity requirements.

All positive hybridoma clones are sequenced at an early stage to eliminate unnecessary screening of redundant clones, accelerate conversion to recombinant Ab and mitigate the risk of any hybridoma instability. This allows informed prioritization of clone testing and ensures customers can test a broad range of sequence-unique clones.


Peripheral Blood B-cell cloning

The second arm of our RabMAb discovery platform interrogates circulating, high-secreting B cells, likely plasma cells.  This technology gives customers the opportunity to find high-affinity antibodies, faster; these antibodies have been selected by the animal for increased expression.  These cells can be isolated early in immunization, and multiple samples can be safely collected at different time points throughout the immunization schedule.

Following isolation of peripheral blood cells and proprietary culture techniques, supernatants from antibody-secreting cells are screened for positive target binding.  Oligoclonal molecular material is recovered from the cultures and subjected to the molecular subcloning steps to generate recombinant monoclonal antibodies.


Next Generation Sequencing

Our proprietary NGS platform builds upon the diversity of the rabbit immune system and enables us to mine the breadth and depth of the immune response, providing a bioinformatic blueprint of target-reactive antibodies. Access to the full immune repertoire yields many thousands of unique antibody sequences, providing maximum diversity in epitope recognition and offering the opportunity to find rare antibody clones with excellent binding properties. Key to the NGS strategy is our single-cell sequencing method allowing for identification of natively-paired light and heavy chains as well as our bioinformatics platform, designed explicitly for rational selection of binders that will be most beneficial to the application. Additionally, NGS is well-suited to be combined with RabMAb discovery approaches to allow for lineage mining for binder discovery and optimization on especially challenging targets or needs.


AxioMx

When antibodies to a target can’t be generated through an in vivo approach, for example, the target is toxic or non-immunogenic, or when a customer needs an antibody exceptionally quick, we can use our phage display technology.

AxioMx phage display represents a rapid approach to antibody discovery, with early indications of success within just 4–6 weeks, and full understanding after 10–14 weeks. This is enabled by our proprietary “Rapid Liquid Screening” technology. We maximize the opportunities of finding suitable clones by using high-diversity M13 phagemid libraries of human antibody single-chain variable fragments (scFvs) followed by a proprietary affinity maturation technology, which increases the efficiency of mutant library generation, thus maximizing diversity. This platform is applied to select and prioritize only the highest specificity binders using both negative depletion of libraries and competition during phage binding.  The result is a recombinant antibody created using animal-free technology.

How do you select antibodies that meet my specific requirements?

In addition to selecting the optimum antibody discovery platform (or combination of platforms), tailor-made assay cascades are designed for each custom antibody discovery project. These cascades are based on the specific requirements of the customer in terms of selectivity, sensitivity and application performance. The cascade sets out the techniques and application-specific tests that will be applied at various stages of the antibody discovery process to find the best binders early in the screening process. The more extensive and well-designed the cascade, the higher the success rate of finding and developing the best antibody.

The process is like a funnel where we narrow down the possible binders. At each step, we apply more precise tests, but we always ask: is this the right test, or is there something else we should do? We progress all our projects in a results-driven fashion and can re-design at any stage of the process to maximize success. We are constantly innovating and finding better ways to predict performance in the final application.

Can you give examples of the types of assays/ testing that can be performed during a custom project?

Assay capabilities

(including but not limited to)

  • Homogeneous high throughput biochemical binding assays (TR-FRET)
  • High throughput pair finding
  • LOQ/LOD determination
  • Matrix Tolerance Assay
  • Ligand/receptor neutralization
  • Functional assays
  • Cell-based binding assays
  • Reporter assays and competition assays
  • Internalization assays
  • ADCC reporter assay


Application-specific screening

  • ELISA
  • Western Blot
  • IHC
  • ICC
  • Flow cytometry

How are your antibodies produced?

All of our custom antibodies are sequenced, cloned, and produced in a recombinant system.  This ensures a high yield of antibody, batch-to-batch consistency, and sustainable long-term supply.

What can your antibodies be used for?

We can produce antibodies for ultimate use in research diagnostics* and as therapeutics*.

In the IVD space, agreements are in place with several of the leading IVD manufacturers, with efficient commercial channels that allow Abcam to act as single point of contact to take reagents from research grade onto multiple IVD platforms.

See here for more details on our partnership strategy.

What quality systems do you have in place?

Central to managing and improving quality is our use of a quality management system that both helps us record and monitor appropriate data and control changes in our manufacturing processes.

Our RabMAb antibody manufacturing site is ISO 9001:2015 certified and all other sites follow the same guidelines.

Our In Vitro Diagnostics (IVD) group is registered with the USFDA and operates under USFDA Quality System Regulation (21 CFR Part 820) and in compliance with ISO 13485:2016. Also, this facility and its quality system have a Medical Device Manufacturing License granted by the California Department of Public Health (CDPH).

What is the timeframe for developing a custom antibody?

The timelines for custom antibody development are variable – they are dependent on the chosen platform, the complexity of the screening cascades, and any application-specific testing requirements.

Projects are structured in a modular fashion, and each module represents an individual invoicing point. At the end of each module, data will be available for review before initiating the next phase of the project. Projects can be modified, paused or terminated at each modular break-point, enabling flexible decision making and helping to minimize time and cost risks. 

The approximate timeframes for antibody development (including delivery of purified antibody) are as follows:

  • RabMAb: 5–8 months (indications of success from 4 months)
  • NGS: 6–9 months (indications of success from 4 months)
  • AxioMx: 2.5–6 months (indications of success from 6 weeks)

What kinds of antigens/immunogens can you use?

We routinely work with a wide variety of immunogens including

  • Recombinant protein
  • Peptide (+/- post-translational modification)
  • Over-expressing cell lines
  • Small molecules (such as nucleotides, organic chemicals, eg PEG)
  • Toxins
  • Polysaccharides

How much does it cost to develop a custom antibody?

The costs for the generation of the custom-antibody will depend on the options chosen.  Abcam offers a fully flexible and data-lead discovery program, with industry experts to support the development of the most appropriate project strategy for your target and antibody requirements.

Our experienced team is happy to review your needs and generate a proposal document outlining suggested strategies alongside estimated costs on a case-by-case basis. We may also be able to design a project to suit your specific budgetary constraints if this is a vital issue.

Will the antibody be sold on Abcam’s catalog?

We offer flexible commercial arrangements on projects and will structure a deal to suit your needs. If feasible, we like to provide the reagents for wider use in the research community. However, we’re also able to offer exclusive use. For some customers, it can be beneficial to have the reagent validated by our customers around the world.

Do you support GMP production?

We can support antibody and assay development to a GMP-ready position and then work with you to transfer your antibody to a partner for GMP manufacture.

Can you support the engineering of the antibody?

Following discovery, if appropriate, we can support a variety of antibody engineering options, including

  • Fc switches
  • Removal of manufacturing sequence liabilities
  • Enhancements to Tm

Can antibodies be humanized for therapeutic purposes?

Several RabMAb antibodies have been successfully humanized for therapeutic use. We can support humanization through working with a partner company and will be happy to discuss this in more detail on a case by case basis.


*Subject to commercial terms and conditions. Please get in touch with partnerships@abcam.com for more information.




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