With more than 14 years’ experience in the antibody development industry, ProteoGenix provides efficient and accurate antibody humanization service based on an enhanced CDR grafting technique. We are committed to collaborate with our customers to help them meet their specific needs; our expertise ensures the success of your project.

Alternatively, our phage display platform can be used to screen antibodies from human libraries to avoid the need to humanize the antibodies.

 

checkFundamental step in the advancement of drug discovery and development

checkPotential to improve biophysical properties of monoclonal antibodies while keeping their affinity and specificity

checkImprove affinity and production yields of the antibody

 

NEED MORE INFORMATION?

  • Your dedicated account manager
    will contact you within 24 hours

CONTACT US


Monoclonal antibody humanization service process

 

Variable regions sequences information are generated by Reverse Transcription of total RNA extraction obtained from hybridoma cell line provided by the customer. Variable regions of the heavy (VH) and light chains (VL) are amplified by PCR and cloned into shuttle vector for sequencing. A total of 5 independent clones are sequenced for each variable chain. Sequences of the hybridoma are determined from the sequencing results of the VH and VL.

A chimeric construct is designed and expressed by combination of mouse VH and VL variable regions with human IgG1 constant regions in order to confirm binding and biological function related to the parental mouse hybridoma.
Antibody sequences are humanized by grafting the three CDRs from the light chain variable region (VL) into human VL germlines which are as homologous as possible to the mouse antibody VL.

Similarly, the three CDRs from the heavy chain variable region (VH) are grafted into human VH germlines which are as homologous as possible to the mouse antibody VH. In addition, because different framework context might bring added value to the resulting antibody, CDRs are grafted into human VH and VL germlines which are well-known to exhibit good overall biophysical properties even if they are less homologous.

Human antibody structure and types of therapeutic monoclonal antibody : mouse, chimeric, humanized and human

Human antibody structure and representation of the distinctions between each type of antibody : Mouse, Chimeric, Humanized and Human.

 

Based on information on the structure of immunoglobulin variable regions, back mutations are performed in the framework regions in which few residues of mouse variable regions are identified as having key roles in maintaining the CDRs in the right conformation or in VH/VL packing. In subsequent versions some of these framework mouse residues can be mutated to the human germline counterpart residue in order to test its real impact on antigen binding or biophysical properties of the molecule.

⇒ A total of 9-18 VH/VL combinations are generated between the CDR-grafted VH, the CDR-grafted VL, and the chimeric versions of both VH and VL.

For more information or to get a quote for Monoclonal Antibody Humanization or Phage display services including the use of human naïve libraries, don’t hesitate to contact us.

Monoclonal antibody humanization service to meet your expectations

 

Proof of concept quantities of each recombinant humanized antibody are produced and evaluated for binding / biological activity / biophysical properties compared to the chimeric version of parental mouse hybridoma.

Following successful testing, a single candidate is selected for further humanization and/or CDRs sequence optimization if required.

Note that for therapeutic or commercial purpose, additional QC can be performed such as HPLC Analysis of Antibody Aggregates and DSC Analysis (Thermostability Analysis by Differential Scanning Calorimetry).