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Microbes & Immunity Anti-mouse CXCR5 monoclonal antibody

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Figure 1. The development of anti-mCXCR5 mAbs. (A) Sprague-Dawley rats were immunized with CHO/mCXCR5 cells. (B) The splenocytes were fused
with P3U1 myeloma cells. (C) Hybridomas producing anti-mCXCR5 mAb were screened using ELISA (first screening) and flow cytometry with CHO-K1
and CHO/mCXCR5 cells (second screening). (D) After limiting dilution, the anti-mCXCR5 mAb (Cx Mab-3) was successfully established.
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Abbreviations: KLH: Keyhole limpet hemocyanin; CHO-K1: Chinese hamster ovary (CHO)-K1; ELISA: Enzyme-linked immunosorbent assay.
screenings, anti-mCXCR5 mAb (Cx Mab-3; rat IgG , CHO-K1 cells. Cx Mab-3 recognized CHO/mCXCR5 cells
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2b
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kappa) was successfully established (Figure 1D). in a dose-dependent manner at 10, 1, 0.1, and 0.01 μg/mL
(Figure 2A). Parental CHO-K1 cells were not recognized
3.2. Flow cytometry analysis using Cx Mab-3 even at 10 μg/mL of any mAbs (Figure 2B). The reactivity
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To assess the reactivity of Cx Mab-3 and L138D7, we of Cx Mab-3 was also observed in LN229/mCXCR5 cells
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conducted flow cytometry analysis on CHO/mCXCR5 and (Figure 3). The sensitivity against CHO/mCXCR5 or
Volume 2 Issue 1 (2025) 104 doi: 10.36922/mi.5664