Development of Antibody Humanization Production Technology-Special topic on antibody modification-Tekbiotech-Yeast Display Service,Phage display technology

Development of Antibody Humanization Production Technology

The monoclonal antibody production technology was invented by Kohler and Milstein in 1975. It can be used not only in basic science, but also in the fields of drugs and biosensors, and has great significance in medicine. Among them, mouse monoclonal antibodies do not contain human gene components and can be recognized by the human immune system. Therefore, they are easy to cause human rejection immune response when used to treat diseases. In order to solve this problem, antibody humanization technology has been developed. The world's first therapeutic antibody was discovered in 1986 and was used to prevent kidney transplant rejection. Since then, many antibody drugs have been found to be used to treat diseases, such as asthma.

At present, the process of antibody humanization has gone through human-mouse chimeric antibodies, humanized antibodies and fully humanized antibodies. The humanized antibody platform can be used for therapeutic antibody discovery and antibody drug discovery.

The development of antibody humanization-tekbiotech.png

Figure 1 The development of antibody humanization

1. Human-mouse Chimeric Antibodies

70% of chimeric antibodies are derived from human genes. Compared with mouse antibodies, they cause fewer adverse reactions and have better therapeutic effects. The preparation of human-mouse chimeric antibodies is as follows:

① Chimeric IgG Antibodies

The V region gene of the target antibody in the hybridoma cell is recombined with the C region gene of the human, cloned into a vector, and transferred into the receptor cell for expression.

② Chimeric Fab and F(ab')2 Antibodies

For most antibodies of this type, due to their low affinity and small molecular weight, they are easily filtered by the glomerulus and disappear from the blood, so most of them are not suitable for clinical treatment alone.

2. Humanized Antibodies

Compared with human-mouse chimeric antibodies, the therapeutic effect is better and the side effects are smaller. The preparation of humanized antibodies is as follows:

① CDR Transplantation (Antibody Reconstruction)

On the basis of retaining the mouse CDR sequence, the human FR sequence is cloned to the corresponding part of the antibody.

② SDR Transplantation

The SDR region refers to the amino acid sequence that can contact the antigen in the CDR region, and the SDR is cloned into the framework of the antibody.

③ Framework Region Reconstruction

Framework region reconstruction is based on the screening of FR, including surface remodeling and glycosylation modification. The former is to humanize the amino acid residues, and the latter is to change the glycosylation site.

3. Fully Humanized Antibodies

Fully humanized antibodies can avoid the body's immune response and have higher safety and efficacy in treatment. Therefore, they are the first choice for treating diseases, especially in the treatment of cancer and autoimmune diseases. The preparation method is as follows:

① Transgenic Mouse Technology

Transgenic mouse technology is to transfer the optimized human antibody DNA sequence into the mouse genome, and the secreted antibodies are all encoded by the human antibody DNA sequence.

② Antibody Library Technology

The immune antibody library is established through single-cell cloning technology.

Non-immune antibody libraries are divided into: semi-synthetic antibodies: composed of randomly synthesized CDR sequences; fully synthetic antibody libraries: variable regions are artificially synthesized; natural antibody libraries: antibody genes are derived from lymphocytes.

③ Phage Display Technology

It is to express variable region genes through phages to screen out antibodies with high affinity.

④ Ribosome Display Technology

This technology is to add antigens to the mRNA-protein-ribosome complex for screening.

⑤ mRNA Display Technology

mRNA display technology is based on ribosome display technology, and the screening is more accurate.

⑥ Yeast Display Technology

Yeast display technology locates exogenous genes on the surface of yeast cells for screening.

⑦ Single cell technology

This technology can screen out single cells from the established antibody library based on cell surface marker molecules, and directly obtain the heavy chain and light chain genes of antibodies by RT-PCR after cell enrichment.

TekBiotech has been committed to the field of antibodies for many years, with rich experience and mature technology, and can provide customers with high-quality antibody humanization preparation and modification, fully human antibody production, and mouse antibody humanization services. In addition, TekBiotech can also provide monoclonal antibody production and antibody customization services, including antibody expression and purification, affinity determination, antibody sequencing, etc., to meet customer needs.

Previous:Recombinant Antibodies Introduction Next:Common Methods for Humanizing Antibodies