Selection of VHH Antibody Fragments That Recognize Different A Depositions Using Complex Immune Libraries
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Phage display technology is frequently used to obtain antigen specific binders with predetermined characteristics. Phage display libraries are often constructed from animals immunized with the antigen of interest. An important point of consideration when making immune libraries is the availability of an appropriate antigen sources. When available, often either the amount is not sufficient for immunization or it is expensive to obtain. To overcome this problem, these antigens are typically obtained by over expression in prokaryotic or eukaryotic expression systems. While this could solve the problem of obtaining sufficient quantities of antigen for a reasonable price and effort, correct folding and differences in posttranslational modification could potentially lead to binders that recognize the recombinant, but not the endogenous protein. In addition, selection of binders against specific modifications or structural epitopes could be missed.
In this chapter we describe a particular selection of VHH antibody fragments from phage display libraries that were constructed from llamas immunized with different complex protein samples containing the antigen of interest. We show that this can result in binders that preferentially recognize the target of interest when present in specific structures depending on the antigen source.
