Monoclonal antibodies that recognise filamentous phage: tools for phage display technology.
We generated six hybridoma cell lines that secrete monoclonal antibodies (mAb) which specifically bind filamentous phage coat proteins. Two of these mAb recognise epitopes that include the N terminus of the coat protein III (pIII), while two others are specific for the N terminus of the major coat protein VIII (pVIII). These mAb are valuable tools to study phage assembly and structure. Furthermore, we describe two examples of how these mAb can be exploited in the construction and screening of peptide libraries displayed by the filamentous phase major coat protein. We have used one of these mAb to develop a sensitive ELISA with crude phage supernatants. This assay allows rapid screening of large numbers of clones from random peptide phage libraries. Some of the anti-phage mAb described here can interfere with wild-type phage propagation, while phage carrying modifications in their coat proteins are insensitive to growth inhibition. We have exploited this observation as a tool to favour the growth of phage displaying peptides fused to pVIII, with respect to vector phage.
Dente
L
L
Dipartimento di Biologia, Università di Roma Tor Vergata, Italy.
Cesareni
G
G
Micheli
G
G
Felici
F
F
Folgori
A
A
Luzzago
A
A
Monaci
P
P
Nicosia
A
A
Delmastro
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D017104:Bacteriophage M13 / Q000276:immunology*; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018604:Epitope Mapping; D005260:Female; D015723:Gene Library; D017912:Inovirus / Q000502:physiology* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins