GreenLight uses phage display technology to increase the binding affinity of a starting antibody. A typical increase of between 10 and 100-fold can be achieved. The options of screening for potential immunogenicity and providing an antibody that avoids T cell epitopes can be added to an affinity maturation project. Our extensive experience of bioassay development enables us to routinely isolate antibodies with equivalent binding specificities to the original antibody and with picomolar binding affinities.
Right: Example improvements achieved by GreenLight showing up to 100-fold increase in affinity.
For phage display technology, antibodies are first converted to a scFv format. Specific amino acids within the CDRs of the antibody are then targeted for “hotspot” mutagenesis. In addition, specific framework residues may be changed or randomised to increase library diversity. Libraries of mutated genes are then constructed and cloned for display on the surface of the phage. Phage are positively selected against reducing concentrations of target antigen and screened using a range of bioassays as required. Lead scFvs can then be converted to a whole IgG, if required. Once made, the lead antibodies can be characterised using an extensive suite of binding assays (such as using ELISA), affinity determination using Biacore Surface Plasmon Resonance or in an appropriate biological assay.
Analogous to affinity maturation, display technologies can also be used to evolve other properties of a molecule, for example, increasing specificity of a molecule to a given receptor over a similar related receptor.
Working with GreenLight
Our services are tailored for each project to ensure that the objectives are met or exceeded. Experienced project teams are assigned to each study focusing on progressing projects through to results in the minimum amount of time. Our clients widely regard us as professional and attentive partners who deliver quality results in a timely manner.
For more information or a quote for our antibody and protein engineering services and technologies contact us.