A library of bacteriophage-displayed antibody fragments directed against proteins of the inner ear

Abstract

Bacteriophage display of antibodies provides a method for the generation of immunological reagents against rare and uncharacterized antigens. To ascertain the usefulness of this approach for the characterization of inner-ear proteins, we produced a bacteriophage-displayed antibody-fragment library directed against proteins from the bullfrog's sacculus. This library was probed for bacteriophage that bound to proteins present in a lysate of hair cells, the sensory receptors of the inner ear. The predominant bacteriophage clone after selection expressed an antibody fragment that recognized a single protein in the inner ear. This antigen occurred in both the nonsensory and sensory epithelia of the sacculus. The specificity of the antibody fragment indicates that our bacteriophage-displayed library provides a useful source of immunological tools that should facilitate the identification and biochemical characterization of novel proteins in the inner ear.

Automated Assessment of the DNA-Binding Capacity of a Proteome by In Vitro Selection

“…Selection methods have previously been applied in whole cell lysates. For example, antibody selections have previously been carried out against cell lysates 8, 9 and have yielded antibodies specific for the antigens within the lysate. Nucleic acid selections against cell lysates have also been performed.…”

De novo identification of cell‐type specific antibody–antigen pairs by phage display subtraction

“…in different cell states. Two groups have independently reported the analysis of cell lysates using different approaches both applying traditional selection strategies, although one uses a novel helper phage and high‐throughput screening on membrane immobilized HeLa lysates [13], and the other analysis of cytosolic content from bullfrog inner‐ear hair cells [14].…”