Expression of anti-Z-DNA single chain antibody variable fragment on the filamentous phage surface.
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.
Maranhão
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Departamento de Biologia Celular, Instituto de Biologia, Universidade de Brasília, Brasília, DF, Brasil.
Brígido
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D000595:Amino Acid Sequence; D000596:Amino Acids / Q000502:physiology*; D000906:Antibodies / Q000276:immunology*; D019677:Artificial Gene Fusion / Q000379:methods; D001435:Bacteriophages / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004279:DNA, Viral / Q000276:immunology*; D005784:Gene Amplification; D015723:Gene Library; D005822:Genetic Vectors / Q000276:immunology / Q000378:metabolism; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000737:chemistry; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction
D000596:Amino Acids; D000906:Antibodies; D004279:DNA, Viral; D007128:Immunoglobulin Fragments; D019151:Peptide Library