To maintain a constant supply of universal blood type, or group O red blood cells, has benefit in specialized transfusion condition. In this study, a-galactosidase cDNA was cloned from Catimor coffee bean grown in Hai...To maintain a constant supply of universal blood type, or group O red blood cells, has benefit in specialized transfusion condition. In this study, a-galactosidase cDNA was cloned from Catimor coffee bean grown in Hainan island, China, by the RT-PCR method. We have constructed a vector for a-galactosidase cDNA expression and transferred a-galactosidase cDNA into Pichia pastoris GS115 cells by elec-troporation. The recombinant a-galactosidase (r-a-GalE) was purified by cation ion exchange chromatography. After studying the biochemical characters of r-a-GalE, we have succeeded in converting human erythrocytes from group B to group O. The animal experiment showed that transfusion of enzymetically converted group O red blood cells (ECORBC) was perfectly safe.展开更多
目的了解A、B、AB型全血加入O型全血不同剂量后血型正反鉴定、抗-A、抗-B抗体效价、直接与间接抗人-IgG实验变化。方法完全随机抽取4℃保存24 h A、B、AB型全血分别60 m l,按不同比例加入O型抗-A、抗-B效价1∶64的全血9、12、15、18 m ...目的了解A、B、AB型全血加入O型全血不同剂量后血型正反鉴定、抗-A、抗-B抗体效价、直接与间接抗人-IgG实验变化。方法完全随机抽取4℃保存24 h A、B、AB型全血分别60 m l,按不同比例加入O型抗-A、抗-B效价1∶64的全血9、12、15、18 m l置入100 m l塑料血袋内混匀后,置37℃孵箱,间隔15 m in摇动1次。分别在1、2、4、8、12、24 h留取标本,做血型正反鉴定、抗-A、抗-B抗体效价、直接与间接抗人-IgG实验。结果不同型全血加入不同剂量O型全血,37℃不同时间血型正反鉴定相符;相对应抗-A、抗-B抗体效价逐渐下降到消失;直接与间接抗人-IgG实验的变化,以B型接受O型的反应最小,在接受600 m l直接与间接抗人-IgG均为阴性,800 m l同1 200 m l反应基本一致。A、AB型接受O型后直接与间接抗人-IgG反应基本一致,实验结果均为阳性。结论以B型接受O型的反应最小,在接受600 m l直接与间接抗人-IgG均为阴性。800 m l同1 200 m l反应基本一致,A、AB型直接与间接抗人-IgG均为阳性。原则上最好不输O型全血,从直接抗人-IgG实验分析,只要红细胞上存在不完全抗体,就一定会影响红细胞的寿命与质量。展开更多
基金supported by the State"863"High-tech Project(Grant No.102-09-04-02)the National Key Basic Research and Development Program(Grant No.2002CB713804)the PLA Ten-Five Key Research Program(Grant No.2000252910).
文摘To maintain a constant supply of universal blood type, or group O red blood cells, has benefit in specialized transfusion condition. In this study, a-galactosidase cDNA was cloned from Catimor coffee bean grown in Hainan island, China, by the RT-PCR method. We have constructed a vector for a-galactosidase cDNA expression and transferred a-galactosidase cDNA into Pichia pastoris GS115 cells by elec-troporation. The recombinant a-galactosidase (r-a-GalE) was purified by cation ion exchange chromatography. After studying the biochemical characters of r-a-GalE, we have succeeded in converting human erythrocytes from group B to group O. The animal experiment showed that transfusion of enzymetically converted group O red blood cells (ECORBC) was perfectly safe.
文摘目的了解A、B、AB型全血加入O型全血不同剂量后血型正反鉴定、抗-A、抗-B抗体效价、直接与间接抗人-IgG实验变化。方法完全随机抽取4℃保存24 h A、B、AB型全血分别60 m l,按不同比例加入O型抗-A、抗-B效价1∶64的全血9、12、15、18 m l置入100 m l塑料血袋内混匀后,置37℃孵箱,间隔15 m in摇动1次。分别在1、2、4、8、12、24 h留取标本,做血型正反鉴定、抗-A、抗-B抗体效价、直接与间接抗人-IgG实验。结果不同型全血加入不同剂量O型全血,37℃不同时间血型正反鉴定相符;相对应抗-A、抗-B抗体效价逐渐下降到消失;直接与间接抗人-IgG实验的变化,以B型接受O型的反应最小,在接受600 m l直接与间接抗人-IgG均为阴性,800 m l同1 200 m l反应基本一致。A、AB型接受O型后直接与间接抗人-IgG反应基本一致,实验结果均为阳性。结论以B型接受O型的反应最小,在接受600 m l直接与间接抗人-IgG均为阴性。800 m l同1 200 m l反应基本一致,A、AB型直接与间接抗人-IgG均为阳性。原则上最好不输O型全血,从直接抗人-IgG实验分析,只要红细胞上存在不完全抗体,就一定会影响红细胞的寿命与质量。
