Phage Display
1) Basic principle behind Antibody-Phage Display
In antibody phage display, the gene encoding for an antibody is inserted into the phage coat gene of a bacteriophage. In this way, the bacteriophage is expected to express said antibody on its surface while containing the antibody’s gene in the phage’s genetic material. Because the antibody is now expressed on the surface of the phage, it is free to interact with other molecules (immobilized antigens). The antibody-displaying bacteriophage are screened using immobilized antigens. The antigen-binding bacteriophage can then be used to create antibody libraries, for further studies on the antibody and its possible uses in drug discovery.
Figure 1. Overview of the steps involved in the generation of human antibody Fab libraries. Messenger RNA is isolated from peripheral blood of donors and copied into cDNA. This material is then used as template for PCR amplification of immunoglobulin gene fragments, which in turn are cloned and expressed as functional Fab molecules in fusion with a phage surface protein (pIII) as mentioned in the text. The entire mixture of phage displayed Fab molecules are then submitted to affinity purification methods to obtain specific binders to a selected “antigen.”
2) Applications of Antibody-Phage Display
Drug Discovery/Therapies
Research (Immune diseases, Cancer)
Creating Antibody Libraries
Producing Antibody Libraries
Works Cited
(n.d.). Image Source, Figure 1: Retrieved June 20, 2014, from Springer Link: http://link.springer.com/protocol/10.1385%2F1-59259-334-8%3A161 (2003). Methods and Protocols III: Human Recombinant Fab Antibodies with T-Cell
Receptor-Like Specificities Generated from Phage Display Libraries. In J.
Engberg, Y. A. F., R. Claussen, L. B. Jensen, P. Sorensen, P. Kopps, et al.,
Recombinant Antibodies for Cancer Therapy (pp. 161-177). Humana Press.
In antibody phage display, the gene encoding for an antibody is inserted into the phage coat gene of a bacteriophage. In this way, the bacteriophage is expected to express said antibody on its surface while containing the antibody’s gene in the phage’s genetic material. Because the antibody is now expressed on the surface of the phage, it is free to interact with other molecules (immobilized antigens). The antibody-displaying bacteriophage are screened using immobilized antigens. The antigen-binding bacteriophage can then be used to create antibody libraries, for further studies on the antibody and its possible uses in drug discovery.
Figure 1. Overview of the steps involved in the generation of human antibody Fab libraries. Messenger RNA is isolated from peripheral blood of donors and copied into cDNA. This material is then used as template for PCR amplification of immunoglobulin gene fragments, which in turn are cloned and expressed as functional Fab molecules in fusion with a phage surface protein (pIII) as mentioned in the text. The entire mixture of phage displayed Fab molecules are then submitted to affinity purification methods to obtain specific binders to a selected “antigen.”
2) Applications of Antibody-Phage Display
Drug Discovery/Therapies
Research (Immune diseases, Cancer)
Creating Antibody Libraries
Producing Antibody Libraries
Works Cited
(n.d.). Image Source, Figure 1: Retrieved June 20, 2014, from Springer Link: http://link.springer.com/protocol/10.1385%2F1-59259-334-8%3A161 (2003). Methods and Protocols III: Human Recombinant Fab Antibodies with T-Cell
Receptor-Like Specificities Generated from Phage Display Libraries. In J.
Engberg, Y. A. F., R. Claussen, L. B. Jensen, P. Sorensen, P. Kopps, et al.,
Recombinant Antibodies for Cancer Therapy (pp. 161-177). Humana Press.
Cited: (n.d.). Image Source, Figure 1: Retrieved June 20, 2014, from Springer Link: http://link.springer.com/protocol/10.1385%2F1-59259-334-8%3A161 (2003). Methods and Protocols III: Human Recombinant Fab Antibodies with T-Cell Receptor-Like Specificities Generated from Phage Display Libraries. In J. Engberg, Y. A. F., R. Claussen, L. B. Jensen, P. Sorensen, P. Kopps, et al., Recombinant Antibodies for Cancer Therapy (pp. 161-177). Humana Press.