Soluble expression, purification, and identification of scFv against carbofuran

The lab constructed anti-carbofuran sc Fv gene, for further study of the nature of anti-carbofuran sc Fv, two kinds of anti-carbofuran sc Fv soluble expression vector p ET22b ( +) -C-6His-CBF-sc Fv and p ET22b ( +) -N-6His-CBF-sc Fv were constructed.N-6His-CBF-sc Fv was better than C-6His-CBF-sc Fv for separation. The conditions of soluble expression and purification effect factors about N-6His-CBF-sc Fv were optimized.The results showed that soluble expression was the best when LB medium was used, the expression temperature was 18 ℃, the expression time was about 16 hours and the concentration of IPTG was 1 mmol/L.Ni Sepharose HP and Iex Cap SP HP were used separate N-6His-CBF-sc Fv.The purity of the purified N-6His-CBF-sc Fv was 90%.Identified by ic ELISA, anti-carbofuran sc Fv maintains antigen binding activity, IC₅₀ value was 63.80 ng/m L, which could be used for the further study.