生物素标记人免疫球蛋白IgG重链的效果评估

目的对生物素标记的人免疫球蛋白IgG重链的效果进行评估和研究。方法将生物素与人免疫球蛋白IgG重链以各种不同的比例进行反应,然后让连接有抗人免疫球蛋白IgG重链抗体的微球颗粒与其反应,应用Luminex200仪器来测定其反应结果,最终来评估生物素与待标记的人免疫球蛋白IgG重链反应的效价。结果生物素与人免疫球蛋白IgG重链的重量比例适当时,微球颗粒上的平均荧光强度(MFI值)最强,而其他比例时,微球颗粒上的平均荧光强度呈下降趋势。结论生物素标记人免疫球蛋白IgG重链应有适当的比例,使被标记的生物素尽可能的多,同时又避免过多的生物素造成空间位阻影响。

To evaluate the efficiency of biotinylation of human immunoglobulin IgG heavy chain

【Objective】To measure the extent of biotin linked to the human immunoglobulin IgG heavy chain.【Methods】Biotin is coupled to heavy chains at different ratios and the efficiency of binding is determined by capturing the labeled heavy chain onto the beads coated with specific antibodies.The intensity of the signal is proportional to the amount of biotin attached to the heavy chain as measured by Luminex 200.【Results】An optimum ratio was reflected by the highest fluorescence.With other ratios there was a decline in the signal.【Conclusion】Optimum biotinylation accounts for the balance between correct ratio and availability of the heavy chain for the antibody.In case where the biotin exceeds,the signal is lower due to steric hindrance at the binding site.