Abstract
We describe the expression of an anti-Z-DNA single chain variable region antibody fragment (scFv) on a filamentous phage surface. Four vectors for phage display were constructed. Two of them are able to display multiple copies of the antibody fragment, and the others can be used to make monovalent libraries. The vectors use different promoter/leader sequences to direct the expression of the fused proteins. All were able to promote the assembly of fusion virion particles. In this paper we also show the affinity selection (biopanning) of those phage-antibodies based on the capacity of their products to recognize the antigen. We used biotinylated Z-DNA and the selection was performed in a solution phase fashion. The data presented here indicate that these vectors can be further used to construct anti-nucleic acid antibody fragment libraries that can be used to study the basis of nucleic acid-protein interaction and its role in autoimmunity mechanisms.
Highlights
Anti-Z-DNA antibodies are found in serum of patients with autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis [1]
We describe the expression of an anti-Z-DNA single chain variable region antibody fragment on a filamentous phage surface
The data presented here indicate that these vectors can be further used to construct anti-nucleic acid antibody fragment libraries that can be used to study the basis of nucleic acid-protein interaction and its role in autoimmunity mechanisms
Summary
Anti-Z-DNA antibodies are found in serum of patients with autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis [1]. The PCR product was digested with both restriction endonucleases and cloned into pIg 16, a vector that codes for the scFv fragment of the anti-Z-DNA Z22 antibody [17].
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