公开(公告)号：US07078179B2

公开(公告)日：2006-07-18

申请号：US10838750

申请日：2004-05-04

申请人： Teresa Di Colandrea ,  Cherisa Meyer ,  Won-Bin Young ,  James N. Higginbotham ,  Mario Mautino ,  Charles J. Link, Jr.

发明人： Teresa Di Colandrea ,  Cherisa Meyer ,  Won-Bin Young ,  James N. Higginbotham ,  Mario Mautino ,  Charles J. Link, Jr.

IPC分类号： G01N33/567 ,  G01N33/53 ,  C12Q1/68 ,  C12N15/63 ,  C12N15/86

CPC分类号： G01N33/5023 ,  C07K16/40 ,  C07K2317/11 ,  C07K2317/23 ,  C12N9/1051 ,  C12N2799/027 ,  G01N2333/91102

摘要： A method is described for the rapid identification and isolation of cells based on the presence or absence of an ectopically-expressed N-acetyllactosaminide 3-α Galactosyltransferase (αGT) enzyme for the production of αGalactosyl-(1,3)Galactosyl (αGal) epitopes on the surface of αGal-negative cells. These cells which are genetically modified to express the αGT enzyme and αGal epitopes on glycosylated lipids and proteins of the cell surface are then labeled via an antibody composition which recognizes and binds the αGal epitopes on the cell surface. Cells labeled with the anti-αGal antibody can be isolated by sorting via fluorescence activated cell sorting (FACS), or by magnetic panning techniques. This method is suitable for the rapid positive or negative selection of αGal-positive cells from within a population of αGal-negative cells without the need to expose cells to antibiotics for any period of time. In addition, the specification provides a method for the production and purification of anti-αGal antibodies from chicken egg yolk.

摘要翻译： 描述了用于快速鉴定和分离细胞的方法，所述方法基于是否存在用于产生α-半乳糖基 - （1,3）半乳糖基（αGal）表位的异位表达的N-乙酰乳糖苷酶3-α半乳糖转移酶（αGT） 在αGal阴性细胞的表面。 然后通过识别和结合细胞表面上的αGal表位的抗体组合物来标记经遗传修饰以表达糖基化脂质和细胞表面蛋白质上的αGT表位的这些细胞。 用抗αGal抗体标记的细胞可以通过荧光激活细胞分选（FACS）进行分选，或通过磁性淘选技术分离。 该方法适用于从αGal阴性细胞群体内快速阳性或阴性选择αGal阳性细胞，无需在任何时间段内将细胞暴露于抗生素。 此外，本说明书提供了从鸡蛋黄制备和纯化抗αGal抗体的方法。

公开(公告)号：US20050250107A1

公开(公告)日：2005-11-10

申请号：US10838750

申请日：2004-05-04

申请人： Teresa Di Colandrea ,  Cherisa Meyer ,  Won-Bin Young ,  James Higginbotham ,  Mario Mautino ,  Charles Link

发明人： Teresa Di Colandrea ,  Cherisa Meyer ,  Won-Bin Young ,  James Higginbotham ,  Mario Mautino ,  Charles Link

IPC分类号： C07K16/40 ,  C12N9/10 ,  C12N15/85 ,  C12P21/04 ,  C12Q1/68 ,  G01N33/50 ,  G01N33/53 ,  G01N33/567

CPC分类号： G01N33/5023 ,  C07K16/40 ,  C07K2317/11 ,  C07K2317/23 ,  C12N9/1051 ,  C12N2799/027 ,  G01N2333/91102

摘要： A method is described for the rapid identification and isolation of cells based on the presence or absence of an ectopically-expressed N-acetyllactosaminide 3-α Galactosyltransferase (αGT) enzyme for the production of αGalactosyl-(1,3)Galactosyl (αGal) epitopes on the surface of αGal-negative cells. These cells which are genetically modified to express the αGT enzyme and αGal epitopes on glycosylated lipids and proteins of the cell surface are then labeled via an antibody composition which recognizes and binds the αGal epitopes on the cell surface. Cells labeled with the anti-αGal antibody can be isolated by sorting via fluorescence activated cell sorting (FACS), or by magnetic panning techniques. This method is suitable for the rapid positive or negative selection of αGal-positive cells from within a population of αGal-negative cells without the need to expose cells to antibiotics for any period of time. In addition, the specification provides a method for the production and purification of anti-αGal antibodies from chicken egg yolk.

摘要翻译： 描述了用于快速鉴定和分离细胞的方法，所述方法基于是否存在用于产生α-半乳糖基 - （1,3）半乳糖基（αGal）表位的异位表达的N-乙酰乳糖苷酶3-α半乳糖转移酶（αGT） 在αGal阴性细胞的表面。 然后通过识别和结合细胞表面上的αGal表位的抗体组合物来标记经遗传修饰以表达糖基化脂质和细胞表面蛋白质上的αGT表位的这些细胞。 用抗αGal抗体标记的细胞可以通过荧光激活细胞分选（FACS）进行分选，或通过磁性淘选技术分离。 该方法适用于从αGal阴性细胞群体内快速阳性或阴性选择αGal阳性细胞，无需在任何时间段内将细胞暴露于抗生素。 此外，本说明书提供了从鸡蛋黄制备和纯化抗αGal抗体的方法。