Purification and characterization of monoclonal IgG antibodies recognizing Ebola virus glycoprotein

Han, Baek-Sang; Jang, Ho-Young; Racine, Trina; Qiu, Xiangguo; Sin, Jeong-Im

MAbs have been broadly utilized as medicine, as well as diagnostic tools for virus infections. MAbs are generated by hybridoma technology where mice are injected with an immunogenic antigen and then the splenocytes are utilized to fuse with myeloma cells [9] . In this regard, antigens of high purity and with native protein folding are a pre-requisite for generating neutralizing antibodies that can target a native form of viral surface proteins. DNA vaccines are thought to be useful at generating the native from of viral antigens as they tend to express their proteins in cells in a manner similar to real viral infection. However, DNA vaccines are less effective at producing IgG-secreting hybridomas, possibly due to the presence of un-methylated CpG sequences in the DNA vaccines that stimulate polyclonal B cell activation [10] . In the Lee et al. (2017) study [10] , use of protein boosting immunization was effective at reversing the preference for production of IgM-secreting hybridomas over IgG-secreting hybridomas. Moreover, multiple immunizations with GP after GP DNA vaccination might be useful at increasing the chance of generating antibodies with higher antigen binding affinity. In this context, it is highly likely that an appropriate application of GP DNA and protein vaccines may generate hybridoma clones that secrete IgG with higher binding affinity to the native form of the antigen.