Comparison of two kinds of protein determination methods for fusion protein Fv-LDP in the recombinant strain
GAO Rui-juan
ZHAO Chun-yan
WANG Yu-feng
LIAN Run-mei
LI Dian-dong
Abstract：Objective:To determine the content of fusion protein Fv-LDP produced by Escherichia coli expressing the single-chain Fv antibody against anti-type Ⅳ collagenase and lidamycin apoprotein (LDP). Methods:The recombinant strain expressing fusion protein Fv-LDP was cultured in shaking flask, and then fusion protein Fv-LDP was purified by the immobilized metal affinity chromatography. Next, the purity of fusion protein Fv-LDP was evaluated by HPLC following the renaturation by step-wise dialysis. The colorimetric method was used to set out the concentration of the recombinant strain. After the analysis of the protein bands by SDS-PAGE, the integrated densi-tometric scanning was performed to determine the expression percentage and the content of fusion protein Fv-LDP. Results:A calibrated linear curve of the concentration of fusion protein Fv-LDP versus the integrated density value was shown within 88.7～1 064 mg· L~(-1)(r=0.998 8). The average recovery rate was 95.18% with the RSD of 2.09% (n=8). Conclusion:This SDS-PAGE-Spot Denso method is more simple, reproducible and sensitive, which can be applied in controlling the production level of fusion protein Fv-LDP during its fermentation process.
Keywords：single-chain Fv antibodylidamycin apoproteinintegrated densitometric scanningcontentfermentation
Publication Date：2010-01-01
Online Publishing Date：2026-08-14(First online date of this platform, not the publication date of the document)
Pages：5( 280-283,331 )