Introduction to Human mAb Identification by Yeast Display

Although phage display technology plays an important role in the antibody discovery process, yeast display technology has some unique advantages in the antibody discovery process. For example, yeast display technology can be better used for antibody discovery projects in which peripheral blood mononuclear cells undergo secondary sorting after animal immunization as well as to avoid missing disulfide bond formation. By setting up the yeast display library construction program and the yeast display library screening program, antibody discovery, screening, identification, and affinity maturation can be achieved quickly. Based on our rich experience and advanced yeast display platform, Creative Biolabs provides comprehensive services to support human monoclonal antibody identification.

Human Monoclonal Antibody Identification

Yeast display technology plays an important role in antibody and protein engineering and thus greatly contributes to the progress of research and development in the field of biopharmaceuticals. For example, by combining with fluorescence activated cell sorting technology, yeast display technology can be used to screen high affinity target antibodies conveniently. In protein engineering, yeast display-based T cell receptor engineering is widely used in T cell receptor antibody drug development and T cell receptor modification for cell therapy. There are already many antibody drugs developed based on yeast display technology that have been marketed for the treatment of clinical diseases. The main advantages of the yeast display system as a eukaryotic display system over other display systems are its efficient post-translational modification mechanism and better adaptation to synthesize complex mammalian proteins with multiple disulfide bonds.

Yeast display antibody library construction begins with cloning a library of heterologous immunoglobulin genes into a yeast display vector and expressing the cloned genes in fusion with the Aga2p protein. Most antibody forms used in yeast display systems are single chain antibodies. However, other human monoclonal antibody forms, including Fabs, single domain antibodies, and intact IgG1, can also be displayed on the yeast surface. Insertion of antibody-encoding genes into yeast display vectors produces libraries of 10⁷ to 10⁹.

In addition to the important applications of yeast display technology in the above fields, it also plays an important role in the field of antibody screening and characterization. In general, antibody screening based on yeast antibody display technology involves the fusion expression of an antibody sequence variable region with the lectin Aga2p, which binds the Aga2p protein subunit to the Aga1p protein subunit immobilized on the yeast cell wall via two disulfide bonds. Magnetically activated cell sorting (MACS) and fluorescence activated cell sorting can be used in the isolation of antibodies from yeast display systems. This eliminates flow cytometry detection rate limitations, increases flow cytometry selectivity, and allows non-binding monoclonal antibody fragments to be removed by MACS. Yeast cells selected with the MACS system labeled with a c-Myc tag can be used to detect ScFv or Fab expression. A more controlled and quantitative selection process is then combined with fluorescence activated cell sorting to screen for and characterize target antigen-specific antibodies.

Creative Biolabs has a wealth of knowledge and experience in yeast display. We would be happy to share with you our knowledge and experience related to human monoclonal antibody identification.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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