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. 2016 Apr 20;7:509. doi: 10.3389/fmicb.2016.00509

FIGURE 1.

FIGURE 1

Production of mAb using wheat germ cell-free synthesized nucleocapsid antigen. (A) Schematic representation of antigen protein production. Recombinant Histidine (His)-tagged N-terminally truncated MERS-NP (122–413) was produced in a wheat germ cell-free system, and then purified using nickel-chelated Sepharose beads. Each protein fraction was analyzed by SDS-PAGE and visualized by CBB staining. Red dot and arrow indicate the target protein. NC, negative control; T, total fraction; S, supernatant; P, precipitate; FT, flow-through; E1–5, elution fractions 1–5. (B) Schematic diagram of hybridoma cells production generating anti-MERS-NP mAb. Purified His-MERS-NP (122–413) was injected into BALB/c mice. After 4 weeks, immunized mouse splenocytes were fused with myeloma cells, and 48 hybridoma cells were established. Of the 48 clones, seven exhibited relatively high reactivity to antigen proteins, as revealed by immunoblotting analysis, and were selected for further investigation. (C) List of selected hybridoma clones producing mAbs.