Construction of single-chain antibodies that bind an overlapping epitope of HIV-1 Nef.
The light and heavy chain variable regions of three mouse hybridoma cell lines (AG11, AE6 and EH1) that produce monoclonal antibodies against an overlapping epitope at the C-terminus of Nef were cloned. Sequence analysis of the light and heavy chain variable regions indicated that clones AG11 and AE6, but not EH1, were highly related. Single-chain antibodies were constructed from the cDNA clones of AG11 and EH1, and subcloned into an eukaryotic expressing vector with the green fluorescent protein as marker for expression. Such intracellular antibodies may provide a way in which to inhibit the function of Nef during HIV-1 infection of cells.
Chang
A H
AH
B.C. Center for Excellence in HIV/AIDS, St. Paul's Hospital, University of British Columbia, Vancouver, Canada.
Hoxie
J A
JA
Cassol
S
S
O'Shaughnessy
M
M
Jirik
F
F
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D015693:Gene Products, nef / Q000276:immunology*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D008164:Luminescent Proteins / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D054311:nef Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D018985:Epitopes, B-Lymphocyte; D015693:Gene Products, nef; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins