Renaturation and Purification of Single Chain Fv Fragment of Monoclonal Antibady Against Haemachrome
重组抗血红素ScFV包涵体蛋白的复性及纯化研究

Most of single chain Fv fragment (ScFv) of monoclonal antibody deposited as insoluble inclusion bodies during expression in E. coli. In order to convert inactive and misfolded inclusion body ScFv into soluble products, the renaturation and purification procedure was developed, recombinant ScFv was redissolved, diluted, and was directly loaded onto a Sephadex G-25 column. The size-exclusion chromatography provided a favorable environment for the proteins to renature, it also spatially constrained partially refolded ScFv from aggregating and diffusing toward each other, promoted renaturation. In the size-exclusion chromatography, the ScFv protein flowed faster and was eluted earlier than the denaturant, this will allow the protein sample to pass through decreasing denaturant concentrations, which promotes ScFv refolding and removes denarurant. During this process, ScFv was simultaneously purified, and at least 150mg/L (with the purity more than 95 %) soluble ScFv was obtained. The strategy provides an economic and novel approach for the production of ScFv from inclusion body proteins.