Characterization and Application of Two Novel Monoclonal Antibodies Against CD154: Epitope and Functional Studies on Cell Membrane CD154 and Studies on the Origin of Soluble Serum CD154
Results Three hybridoma cell lines(1A12、3C11、3B4) producing monoclonal antibodies against the recombination protein HPhA were established by the fusion technique. The three mAb belonged to IgG1、IgM and IgG2b respectively.
Western blotting showed that the protein had a high specificity against mouse-anti-Sj14-3-3 monoclonal antibody and rSj14-3-3 had a promising immune reactivity.
Several indirect plasminogen (Pg) activators are known including streptokinase and the monoclonal antibody IV-Ic, whose mechanism of activation is well studied.
To characterize thermodynamically the activation of Pg by streptokinase (SK) and the monoclonal antibody (mAB) IV-Ic, the activation energies were calculated for various reaction stages.
The sensitivity of enzyme immunoassay based on all monoclonal antibodies was higher compared to analysis based on polyclonal rabbit antibodies (0.1 and 0.4 ng/ml, respectively).
The binding site of MAB F8 is in the immunodominant region of the TPO molecule, in the vicinity of the autoantigenic determinants, whereas the epitope specific for MAB A1 lies outside this location.
Five types of interaction of the AFP-MAB complex with the MAB fixed on NCM were found: 1) complete neutralization; 2) partial neutralization; 3) unidirectional neutralization; 4) enhanced binding; 5) lack of interaction.
Among the resulting monoclonal antibodies, two Ca2+-dependent clones (mAb3 and mAb19) recognizing recoverin were detected by solid-phase immunoenzyme assay.