ABSTRACT
Objective To evaluate the repeatability and stability of double antibody sandwich ELISA kit for micro-quantitative soluble complement receptor 1(sCR1) in human serum and to understand its practical application effect .Methods 50 patients with middle and advanced liver cirrhosis and 50 individuals undergoing physical examination served as the liver disease group and normal control group respectively .The mouse anti-human CD35 monoclone antibody ,rabbit anti-human sCR1 polyclonal antibody and goat anti-rabbit IgG labeled by horseradish peroxidase served as the envelope antibody ,sandwich antibody and detection antiboby .The purified recombinant human sCR1 protein served as the standard substance .The human serum micro-quantitative double antibody sandwich CR1 ELISA kit was established .Then the repeatability and stability tests were performed .Then the sCR1 protein level of two group of serum was detected by this kit .Results The linear range of double antibody sandwich ELISA for detecting human se-rum micro-quantitative sCR1 protein was 15 .60 -250 .00 ng/mL ;the regression equation of sCR1 protein concentration to absor-bance value was Y=112 .10X2 +18 .21X+1 .694(r2 =0 .998);in the repeatability test ,the intra-batch relative standard deviation (RSD) in high and low concentrations of standard substance detection value was 6 .20% and 7 .40% respectively ,the inter-batch RSD was 6 .70% and 7 .90% respectively ;in the stability test ,RSD was not more than 0 .01;the serum sCR1 expression level in the liver disease group was significantly higher than that in the normal control group (P<0 .01) .Conclusion The human serum double antibody sandwich ELISA kit for detecting human sCR1 has wide linear range ,good repeatability ,is easy to be stored and suitable for clini-cal and scientific research detection work .
ABSTRACT
Objective To prepare highly expressed, purified and refolded SCR15-18 of human soluble complement receptor type 1 (sCR1-SCR15-18) protein. Methods The expression of recombinant pET32-sCR1-SCR15-18 in E.coli.. BL21 was induced by IPTG of different concentrations for different time period under different temperatures and the bacteria were split by sonication. The sCR1-SCR15-18 protein was purified by Ni2+-NTA resin affinity chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The sCR1-SCR15-18 protein of high expression, purity and bioactivity was attained. Conclusion The parameters of expression, purification and refolding of sCR1-SCR15-18 protein were optimized, which may pave a way for further studies.
ABSTRACT
Objective:To evaluate the protective effects of adenovirus-mediated gene transfer of soluble complement receptor type 1(sCR1) on acute myocardium ischemia in mice.Methods: Twenty-seven SD rats were subjected to left anterior descending coronary artery(LAD) occlusion(30 min) and reperfusion.In the treatment group(n=14),a mixture of adenovirus carrying sCR1 and LacZ was injected into the ischemic zone(100 ?l,10~(10)pfu)5 min before reperfusion;in the control group,only adenovirus carrying LacZ was injected(n=13).Echocardiography was performed 2 weeks later,followed immediately by pathologic examination.Results: Echocardiographic results of the treatment group were better than those of the control group((P