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作者分别用差速离心法及蔗糖梯度离心法制备胎盘微绒毛膜(PMM),用受体放射分析法研究了PMM的转铁蛋白受体(TfB),结果表明:两种方法制备的PMM均可用于受体分析,但差速离心法更为简便。测定60例产妇PMM的TfR,其数目为3.53±1.98×10~(12)个位点/mg膜蛋白,最大结合容量为6.33±4.21×10~(-12)mol/mg膜蛋白,Kd值为4.95±3.39×10~(-9)mol/L。受体结合反应体系最适实验条件为,膜蛋白浓度每管50μg,标记物浓度每管50 000cpm,孵育30分钟,B/F分离的聚乙二醇浓度为12％(W/V)。对TfR的特性研究表明:TfR与转铁蛋白的结合具有高度亲和力、高度特异性及可饱和性。
The authors used differential centrifugation and sucrose gradient centrifugation to prepare placental microvilli (PMM), respectively. The receptor radiometric assay was used to study the transferrin receptor (TfB) of PMM. The results showed that both PMM Can be used for receptor assays, but differential centrifugation is easier. The TfR of 60 maternal PMMs was determined, which was 3.53 ± 1.98 × 10 ~ (12) sites / mg membrane protein and the maximum binding capacity was 6.33 ± 4.21 × 10 ~ (-12) mol / mg membrane protein. The Kd value 4.95 ± 3.39 × 10 ~ (-9) mol / L. The optimum conditions for the receptor-bound reaction system were as follows: the concentration of membrane protein was 50 μg per tube, the concentration of marker was 50 000 cpm per tube, and the concentration of polyethylene glycol was 12% (W / V). TfR characteristics of the study showed that: TfR and transferrin binding with high affinity, high specificity and saturability.