Affinity Tune-Up & Humanization

When considering antibodies for therapeutic purposes, it is important to be mindful of the immunogenicity of the drug. Antibody humanization can reduce the immunogenicity of monoclonal antibodies derived from other sources, while also improving efficacy, safety due to toxicity, and activation within the immune system.

Antibody humanization works by replacing non-human antibody structures with human ones. This is a key point to increase their similarity to antibody variants produced naturally in humans, and is important to note when using antibodies for therapeutic uses. Depending on your application, ProMab offers an affinity tune up and maturation service to increase antibody function and specificity to your needs. Antibodies derived from other sources than human may also have low or varying affinity, so when paired with our antibody humanization services, we can tune the antibody to the required goal.

Affinity tune-up and maturation goes hand in hand with antibody humanization services. ProMab is backed with nearly 20 years of antibody engineering experience and utilizes state of the art 3D modeling software to ensure we can provide the best service for our clients. We strive to lead the competition for therapeutic antibody discovery and development.


ProMab Biotechnologies Custom Humanization Service: A complete service, from antigen to clinically relevant solution.

At ProMab Biotechnologies, we perform high-throughput screening of our human antibody library using the Octet ForteBio label-free sensitive binding system or SPR (surface plasmon resonance). Clones from the human antibody library are individually tuned, and varying binding activities are generated for preclinical and clinical use. The goal of this high-throughput screening assay is to generate antibodies with no (or low) affinity for antigens in normal cells but with high affinity for cancer cells.

This model shown below is what ProMab uses for many of its proprietary CAR-T technologies, as humanizing the antibody scFv sequence can possibly lead to improve efficacy in the CAR-T setting.


Figure 1. The scheme for antibody humanization for generating CAR-T cells.

To decrease the immunogenicity of monoclonal antibodies derived from rodents, we use a rational approach based on antibody sequence and 3D structure by computer modeling and docking programs. In brief, steps include: hybridoma sequencing, analysis of the scFv by different bioinformatics and computer programs, generating best models and testing these antibodies efficacy.

Alongside ProMab's technology platform, we can ultimately retest these humanized scFvs in a CAR-T/NK setting using various in vitro or in vivo studies.


Figure 2. An illustration of the stepwise process of humanizing a rodent mAb for generating a CAR-T cell with a humanized scFv.

This is done by a group of bioinformaticists and computer modeling professionals with many years of experience in antibody humanization with references (1-3) below.


  1. Alexey Teplyakov, Galina Obmolova, Thomas J. Malia, Jinquan Luo, Salman Muzammil, Raymond Sweet, Juan Carlos Almagro & Gary L. Gilliland. Structural diversity in a human antibody germline library. mAbs. V9, 2016, 1045-63.
  2. Almagro J.C., Teplyakov A., Luo J., Sweet R.W, Kodangattil S, hernandez-Guzman F, Gilliand G.L. Second antibody modeling assessment (AMA-II). Proteins 2014; 82: 1553-1562.
  3. Almagro JC, Fransson J. Humanization of antibodies. Front Biosci., 2008, 1;13:1619-33.

Other Custom Services:

Mouse monoclonal antibodies

Human monoclonal antibodies


Hybridoma Sequencing