Part:BBa_K2622004:Experience

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Applications of BBa_K2622004. scFv merged to membrane proteins, can be used as indicator for dissplay and membrane protein evolution. . ===

Protein is functional, but need additional stabilization to prevent aggregtion and loss of function. ScFv_antiVLY is an antibody against toxin VLY. By binding to VLY, scFv inhibits toxins ability to lyse cells. This was comfirmed by experiment with red blood cells. While working with this system, we saw, that the activity of scFv highly depends on time, which protein spend on ice or in +4 C. If experiment takes place just after expression (we used IVTT system for expression), scFv shows high activity, but if the protein is left for some time in +4 C, there is a possibility, that the activity won't be detected. It is due to insolubility of scFv. Protein shortly aggregates and loses function.

In all experiments, VLY concentration was 3 ng/mL, while scFv concentrations varied.

UNIQ1006dece385edb81-partinfo-00000000-QINU UNIQ1006dece385edb81-partinfo-00000001-QINU