| Publication number |
CLPUB00645 |
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| Authors |
Langton B.C., Sliwkowski M.X., Tran K.V., Knapp S., Keitelmann E., Smith C.M., Wallingford S., Liu H.-L.C., Ralston J.S., Brandis J.W., Coates S.R. |
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| Title |
Development and characterization of monoclonal antibodies to the HTLV-I Tax (P40X) protein. |
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| Citation |
(In) Medical virology 8; de la Maza L.M., Peterson E.M. (eds.); pp.295-295; Plenum Press; New York (1989) |
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| Abstract |
The 40 Kd tax (p40X or tat) protein from HTLV-I, known to exhibit
transcriptional transactivation activity has been cloned and expressed in
E. coli as a trpE fusion protein. This recombinant protein was purified to
homogeneity and used as an immunogen to develop several monoclonal
antibodies (MABs) to tax, of the IgM and IgG2a isotypes. Spleen cells from
immunized Balb/c mice were fused with P3X63Ag8 myeloma cells, screened on
an ELISA assay to the recombinant protein, cloned by limiting dilution and
ascites prepared and purified by HPLC. Selected MAbs were further
characterized by several methods including Western blot,
radioimmunoprecipitation, competition ELISA assay and Geysen epitope
mapping. All of the MAbs recognize the recombinant tax protein and several
of the MAbs also recognize the native tax from a lysate of an HTLV-I
transformed cell line, SLB-1. This binding to native tax can be competed
by the recombinant protein. Competition ELISA assays with patient sera,
known to contain antibody to the tax protein, reveal that the MAbs and
some patient sera may recognize similar epitopes on the protein. In an
attempt to localize binding sites on tax, mapping by Geysen technology has
revealed a few potential epitopes for selected MAbs. Work is in progress
to complete this mapping and determine if these sites are relevant binding
sites for tax reactive antibodies in patient sera. In conclusion, anti-tax
MAbs have been developed which are useful both for the detection and
quantitation of native and recombinant tax. These MAbs are also being used
to localize binding sites on the tax protein.
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| Cell lines |
CVCL_A0AE; 168A51-2 CVCL_A0AF; 168A51-42 CVCL_A0AA; 168B17-46-34 CVCL_A0AB; 168B17-46-50 CVCL_A0AC; 168B17-46-70 CVCL_A0AD; 168B17-46-92 |
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