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    • 21. 发明授权
    • Streptomyces griseoplanus comprising an α-galactosidase for removing immunodominant α-galactose monosaccharides from blood group B or AB reactive cells
    • 包含α-半乳糖苷酶的灰色链霉菌,用于从血型B或AB反应性细胞中除去免疫优势的α-半乳糖单糖
    • US08697411B2
    • 2014-04-15
    • US13205588
    • 2011-08-08
    • Henrik ClausenHumberto de la VegaCheryl HillQiyong Peter Liu
    • Henrik ClausenHumberto de la VegaCheryl HillQiyong Peter Liu
    • C12N9/24
    • C12N5/0641C12N9/2402C12N9/2465C12Q1/34C12R1/465C12Y302/01022C12Y302/01049
    • This invention relates to enzymatic removal of type A and B antigens from blood group A, B, and AB reactive cells in blood products, and thereby converting these to non-A and non-B reactive cells. The invention further relates to using unique α-N-acetylgalactosaminidases and α-galactosidases with superior kinetic properties for removing the immunodominant monosaccharides of the blood group A and B antigens and improved performance in enzymatic conversion of red blood cells. The preferred unique α-N-acetylgalactosaminidases and α-galactosidases exhibit the following characteristics: (i) exclusive, preferred or no less than 10% substrate specificity for the type A and B branched polysaccharide structures relative to measurable activity with simple mono- and disaccharide structures and aglycon derivatives hereof; (ii) optimal performance at neutral pH with blood group oligosaccharides and in enzymatic conversion of cells; and (iii) a favorable kinetic constant Km with mono- and oligosaccharide substrates. The conversion methods of the invention use significantly lower amounts of recombinant glycosidase enzymes than previous and result in complete sero-conversion of all blood group A and B red cells.
    • 本发明涉及从血型A,B,AB型反应性细胞中分离A型和B型抗原,从而将其转化为非A型和非B型细胞。 本发明还涉及使用独特的α-N-乙酰半乳糖胺酶和具有优异动力学性质的α-半乳糖苷酶,用于去除血型A和B抗原的免疫优势单糖,并提高了红细胞酶促转化的表现。 优选的独特的α-N-乙酰半乳糖胺酶和α-半乳糖苷酶具有以下特征:(i)相对于简单的单糖和二糖的可测量的活性,对于A型和B型支链多糖结构,独特的,优选的或不低于10%的底物特异性 结构和糖苷配基衍生物; (ii)在中性pH下用血型寡糖和细胞酶促转化的最佳性能; 和(iii)具有单糖和寡糖底物的有利的动力学常数Km。 本发明的转化方法比以前使用显着更低量的重组糖苷酶,并导致所有血型A和B红细胞的完全血清转化。