Avian leukosis virus is a versatile eukaryotic platform for polypeptide display.
Display technology refers to methods of generating libraries of modularly coded biomolecules and screening them for particular properties. Retroviruses are good candidates to be a eukaryotic viral platform for the display of polypeptides synthesized in eukaryotic cells. Here we demonstrate that avian leukosis virus (ALV) provides an ideal platform for display of nonviral polyaeptides expressed in a eukaryotic cell substrate. Different sizes of polypeptides were genetically fused to the extreme N-terminus of the ALV envelope glycoprotein in an ALV infectious clone containing an alkaline phosphatase reporter gene. The chimeric envelope glycoproteins were efficiently incorporated into virions and were stably displayed on the surface of the virions through multiple virus replication cycles. The foreign polypeptides did not interfere with the attachment and entry functions of the underlying ALV envelope glycoproteins. The displayed polypeptides were fully functional and could efficiently mediate attachment of the recombinant viruses to their respective cognate receptors. This study demonstrates that ALV is an ideal display platform for the generation and selection of libraries of polypeptides where there is a need for expression, folding, and posttranslational modification in the endoplasmic reticulum of eukaryotic cells.
Khare
Pranay D
PD
Molecular Medicine Program, Mayo Clinic Rochester, Rochester, MN 55905, USA.
Russell
Stephen J
SJ
Federspiel
Mark J
MJ
D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics* / Q000378:metabolism; D016466:CHO Cells; D002272:Carcinoembryonic Antigen / Q000276:immunology; D002460:Cell Line; D002645:Chickens; D006224:Cricetinae; D015752:Genes, env; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism*; D011991:Receptors, Virus / Q000502:physiology; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins / Q000096:biosynthesis
D002272:Carcinoembryonic Antigen; D019151:Peptide Library; D010455:Peptides; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins