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| PCR Cloning and Subcloning | Mouse monoclonal IgG were raised using the GenScript MonoExpressTM 106 protocol 107 (Genscript, Piscataway, NJ). Briefly, five mice were immunized with the purified recombinant protein and T-max TM 108 adjuvant. Spleen cells were isolated and fused with 109 myeloma cells and selected hybridomas were screened by ELISA for reactivity with the 110 recombinant protein. Selected positive parental clones were subcloned by limited 111 dilution. | Get A Quote |
Antigen-based immunoassays are currently needed for point-of-care quantification 15 of Loa loa microfilariae (mf). Coupling transcriptomic approaches with bioinformatic 16 analysis, we have identified 11 specific putative proteins (coding mRNAs) with potential utility as biomarkers of patent (mf+ 17 ) L. loa infection. We successfully developed antigen 18 capture immunoassays to quantify 2 (LOAG_14221 and LOAG_15846) of these proteins 19 in individual plasma/sera. Of the 2 quantifiable circulating biomarkers, LOAG_14221 20 showed the highest degree of specificity, particularly with a monoclonal antibody-based immunoassay. Moreover, the levels of LOAG_14221 in L. loa mf+ 21 patients were 22 positively correlated... More
