Improve the affinity of your antibody from five to over a thousandfold thanks to our guaranteed antibody affinity maturation service. At ProteoGenix, we combine our expertise in historical phage display with our renowned knowledge of affinity maturation, helping you pave the way to the clinic! Because we are fully committed to the success of your project, our antibody affinity maturation service includes unique guarantees defined after a preliminary study.

Antibody optimization by affinity maturation

ProteoGenix has developed a high-class affinity maturation service tailored to increase the affinity of your antibody against its target. Besides affinity, cross-reactivity can also be introduced to avoid the use of surrogate antibodies for preclinical development. Many other parameters can be measured and optimized such as:

  • Stability
  • Immunogenicity
  • Chemical liabilities

To do so, site-directed mutagenesis or random mutagenesis is performed on the CDR regions.

  • Site-directed mutagenesis is based on 3D modeling in order to identify key residues involved in the antibody-antigen interaction. Specific mutations on these residues are performed in order to create a large phage display library with low junk variants and high probability of strong affinity enhancement.
  • Random mutagenesis are inserted by the use of error prone PCR. This method allows the construction of large libraries containing 108 starting from 3 to 10 selected antibodies.

Which strategy should you choose
for your affinity maturation project?

As affinity maturation is fairly complex, our experts define the most relevant strategy after a preliminary study. This is the best way to guarantee our customers a safe investment and to include unique result guarantees to your project. This demonstrates our commitment to the success of your antibody affinity maturation.

Our antibody affinity maturation process

Preliminary study by our affinity maturation expert

  • Binding test in relevant application
  • Construction of a phage display screening protocol
  • Definition of a strategy
  • Assessment report including evaluation of your project’s feasibility and chance of success

Design and construction of a mutated phage display library

  • Library design using the mutagenesis strategy previously established
  • Creation of an antibody DNA variant library
  • Creation of the phage display library

Library screening and variants selection

  • Library screening using the custom protocol established in first phase (4 to 6 panning rounds)
  • Validation of the best phage variants in the relevant application
  • Affinity comparison of the best single phages

Sequencing and recombinant expression of the best variants

  • Antibody sequencing
  • Gene synthesis and subcloning in an adapted expression vector
  • Transient expression test in mammalian cells

Antibody tests

  • Antibody characterization depending on customer’s request:
  • KD determination: Biacore (SPR), Xelplex (SPRi), OctetRed
  • Binding analysis: ELISA
  • Stability analysis: DSC
  • Aggregate analysis: SEC-HPLC
  • Other options available
Step Content Timeline Deliverables
Preliminary study
  • Antibody expression on phage surface
  • Binding test
  • Establishment of custom phage display screening protocol
~2 months
  • Detailed report including evaluation of your project’s feasibility and chance of success, description of the project strategy and guarantees provided
Phage display library design and construction
  • Library design
  • Creation of mutated genes library
  • Creation of phage display library
~2 months
  • Detailed report
Library screening and variants selection
  • Library screening
  • Screening and validation of the best phage variants in relevant application
  • Affinity comparison via titer and KD determination
~2 months
  • Detailed report
Sequencing and recombinant antibody expression
  • Antibody sequencing DNA synthesis and sub-cloning into an expression vector
  • Transient expression test in mammalian cells
~2 months
  • Test sample after transient expression test (up to 1mg purified antibody)
  • 1 to 100mg purified antibody depending on customer requirements Detailed report
Antibody characterization
  • ELISA binding analysis
  • SEC-HPLC aggregate analysis
  • DSC stability analysis
  • Antibody KD determination by Biacore, XelPlex or Octet
~1-3 weeks
  • Detailed report

Options available:

When should you envisage antibody affinity maturation?

Affinity maturation is important in diagnostics and biotherapeutics as well as many other applications requiring strong antibody-antigen interactions.

When developing antibodies, many features can be required such as functionality (inhibiting activities for instance), specificity, use in particular applications and of course affinity. But it can be tricky to select an antibody with all these features from the start. It is therefore relevant to first focus on the functionality and application before working on improving the affinity using our high-class phage display platform.

For more information or to get a quote for our phage display services with the use of Affinity Maturation, don’t hesitate to contact us.