Preparation and functional identification of a monoclonal antibody against the recombinant soluble human NKp30 receptor

Generation of a monoclonal antibody recognizing the heavily glycosylated CD45 protein and its application on identifying circulating tumor cells

“…Only 1 cell fusion and 2 cycles of sub-cloning were performed. In contrast, in previous studies [ 4 ] employing peptides or prokaryotic protein as the antigen, 2 or more cell fusions and hybridoma screens were performed. For most laboratories conducting biological research, efficient time and labor management are key factors influencing the decision whether to generate specific mAbs in house.…”

“…The potential impact of glycosylation on antibody recognition has not been comprehensively investigated. Still, some studies [ 4 ] inferred that the mismatch between unglycosylated antigen and glycosylated target protein might cause the fluctuations in antibody binding. The purpose of this study was to provide a reference for biological laboratories demanding appropriate mAbs and facing a similar dilemma: i.e., what kind of antigen should be used (eukaryotically expressed protein, prokaryotically expressed protein, or peptides).…”

“…Moreover, economic considerations have prompted most commercial mAb providers to employ prokaryotic proteins, or peptides, instead of eukaryotically expressed proteins as the antigen. The lack of post-translational modification (for instance, glycosylation) can compromise the consistency and repeatability of studies involving glycosylated proteins [ 4 ]. To avoid these flaws of commercial mAbs, generating mAbs in one’s own laboratory may be considered as a feasible alternative, especially while performing studies on post-translationally modified proteins.…”

TLR2 Limits Development of Hepatocellular Carcinoma by Reducing IL18-Mediated Immunosuppression

“…Liver tissues and hepatocytes were collected for Western blot analysis as previously described (21). IL18, TLR2, and caspase-8 were detected after incubation with the corresponding primary antibodies: anti-IL18, anti-TLR2 (Abcam), and anti-caspase-8 (Cell Signaling Technology), respectively.…”