Figure 1
Figure 1. Constructs and purified protein products from cell-free reactions. (A) Diagram of 38C13 idiotype constructs produced in the cell-free expression system. Construct 1 is an scFv linked to mouse GM-CSF via an Im9 linker. Construct 2 links human GM-CSF to the scFv fragment. The third construct is a fusion of IL-1β9aa to the 38C13 scFv. Construct 4 is the 38C13 scFv alone. Constructs not shown are the C-terminal IL-1β9aa fusion to the 38C13 scFv, the A20 scFv, and the E coli disulfide bond isomerase (DsbC). (B) After 4 hours of protein synthesis, products were enriched from the cell-free reaction supernatant using a nickel affinity column. The partially purified products of constructs were analyzed by SDS-PAGE and stained with Coomassie blue. The arrows indicate the band that corresponds to the desired product. For comparison, unpurified cell-free product is shown (lane U).

Constructs and purified protein products from cell-free reactions. (A) Diagram of 38C13 idiotype constructs produced in the cell-free expression system. Construct 1 is an scFv linked to mouse GM-CSF via an Im9 linker. Construct 2 links human GM-CSF to the scFv fragment. The third construct is a fusion of IL-1β9aa to the 38C13 scFv. Construct 4 is the 38C13 scFv alone. Constructs not shown are the C-terminal IL-1β9aa fusion to the 38C13 scFv, the A20 scFv, and the E coli disulfide bond isomerase (DsbC). (B) After 4 hours of protein synthesis, products were enriched from the cell-free reaction supernatant using a nickel affinity column. The partially purified products of constructs were analyzed by SDS-PAGE and stained with Coomassie blue. The arrows indicate the band that corresponds to the desired product. For comparison, unpurified cell-free product is shown (lane U).

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