Phage Display Library Screening by Affinity Isolation of Antigen-Specific B Cells

Creative Biolabs has developed a strategy for affinity isolation of antigen-specific circulating B cells of interest for subsequent generation of immune antibody phage display libraries. This method overcomes the problem of low yields of monoclonal antibodies of interest in the libraries, which are commonly generated from peripheral blood mononuclear cells (PBMCs) caused by the low abundance of antigen-specific B cells. Based on this strategy, our scientists have significantly increased the abundance of antibody phage particles of interest in the library. We can provide high-quality monoclonal antibodies with the predefined specificity for all our clients.

Starting Materials Commonly Used for Phage Library Construction

Human bone marrow is a major repository for maturated antibody-secreting plasma cells, which produce the majority of the antibodies found in serum. This makes bone marrow an attractive source of RNA for generating human immune antibody libraries to isolate monoclonal antibodies with similar specificity as that found in the donor serum. Unfortunately, bone marrow is not always readily available, because health and ethical concerns often prohibit bone marrow aspiration and leave PBMCs as the only readily available source. Although human immune libraries can be generated from circulating B cells, the low frequency of antigen-specific B cells in the circulation may limit representation of monoclonal antibodies of interest in the resulting library.

Fig.1 Affinity isolation of antigen-specific circulating B cells using antigen-conjugated magnetic beads for subsequent generation of phage display libraries (Ditzel 2009).

Phage Display Library Screening by Affinity Isolation of Antigen-Specific B Cells in Creative Biolabs

Creative Biolabs has developed a preselection strategy to enrich antigen-specific B cells prior to antibody library construction. The preselection of B cells is based on the specificity of the surface Ig receptor and is accomplished using antigen-conjugated magnetic beads. The availability of magnetic beads with different surfaces is suitable for antigen binding, which makes this strategy far more efficacious. We have applied this approach to isolate a panel of target-specific monoclonal antibodies derived from circulating B cells of interest. This strategy can be widely applicable to isolate antibodies against viral antigens, infectious antigens, autoantigens, alloantigens, etc.

Key Advantages Including but Not Limited to

Overcome the problem of low yields of monoclonal antibodies of interest in the libraries.
Libraries are generated from B cells preselected, instead of PBMCs.
Increase the abundance of antibody phage particles of interest.
Diverse antigens can be targets, such as viral antigens, infectious antigens, autoantigens, alloantigens, etc.

Creative Biolabs has accumulated 10 years of experience in therapeutic and diagnostic antibody development. We are proud to introduce this new strategy to all our global clients and facilitate their meaningful antibody related researches. We offer turn-key or ala carte services customized to our client’s needs. Please feel free to contact us for more information.

Reference

Ditzel, H.J. Affinity isolation of antigen-specific circulating B cells for generation of phage display-derived human monoclonal antibodies. Methods Mol Biol. 2009, 562:37-43.

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