Soluble thymic stromal lymphopoietin receptors are absent in murine sera--detection with anti-mTSLPR monoclonal antibodies.
Two monoclonal antibodies, termed nnIE11 and nnIG11, were generated against the murine thymic stromal lymphopoietin receptor, mTSLPR, using traditional hybridoma technology. The antibody-producing hybridoma clones were obtained by fusing P3X63-Ag8.653 myeloma cells with splenocytes from Balb/c mice immunized with anti-FLAG M2 affinity-purified FLAG-tagged mTSLPR from pSVL-mTSLPR-FLAG-transfected COS cells and Ni-NTA-purified his-tagged mTSLPR from recombinant FastBacHisB-mdelta1 baculovirus-infected Sf9 cells. Several monoclonal anti-mTSLPR-specific hybridoma clones were obtained and two of these clones are further characterized here. The generated antibodies could in an immunoblotting identify baculovirus-expressed mTSLPR proteins with a molecular weight corresponding to 50 kDa. Both immunoblotting and ELISA with recombinant mouse TSLPR/Fc chimera as antigen, having only the N-terminal domain of mTSLPR present, indicated that the generated monoclonal antibodies identify the C-terminus of mTSLPR. Although sandwich ELISAs performed with a goat anti-mTSLPR antiserum as capture antibody and nnIE11 as indicator antibody were able to detect mTSLPR in the range of 5 ng/ml, no souble mTSLPR could be observed in serum samples from CBA/H, Balb/c and C57Bl/6 mice.
Rosenqvist
Linda
L
Department of Biology, Abo Akademi University, Turku, Finland.
Andersson
Anne-Marie
AM
Ohls
Sara
S
Boström
Marina
M
Nordström
Tommy
T
Lindqvist
Christer
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D007136:Immunoglobulins; D051379:Mice; D018121:Receptors, Cytokine / Q000097:blood* / Q000276:immunology
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D018121:Receptors, Cytokine; C410591:Tslpr protein, mouse