用钙衍生化羟基磷灰石进行Fab的反向钙亲和纯化。
Reverse calcium affinity purification of Fab with calcium derivatized hydroxyapatite.
作者信息
Gagnon Pete, Cheung Chia-Wei, Yazaki Paul J
机构信息
Validated Biosystems, San Clemente, CA 92672, USA.
出版信息
J Immunol Methods. 2009 Mar 15;342(1-2):115-8. doi: 10.1016/j.jim.2008.11.021. Epub 2009 Jan 14.
Abstract
This study introduces the application of calcium-derivatized hydroxyapatite for purification of Fab. Fab binds to native hydroxyapatite but fails to bind to the calcium derivatized form. IgG, Fc, and most other protein contaminants bind to the calcium form. This supports Fab purification by a simple flow-through method that achieves greater than 95% purity from papain digests and mammalian cell culture supernatants. Alternatively, Fab can be concentrated on native hydroxyapatite then eluted selectively by conversion to the calcium-derivatized form.
摘要
本研究介绍了钙衍生化羟基磷灰石在纯化Fab中的应用。Fab可与天然羟基磷灰石结合，但不能与钙衍生化形式结合。IgG、Fc和大多数其他蛋白质污染物可与钙形式结合。这支持通过简单的流通方法纯化Fab，该方法可从木瓜蛋白酶消化物和哺乳动物细胞培养上清液中获得大于95%的纯度。或者，Fab可在天然羟基磷灰石上浓缩，然后通过转化为钙衍生化形式进行选择性洗脱。
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Biotechnol Bioeng. 2005 Dec 20;92(6):665-73. doi: 10.1002/bit.20729.
2
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J Chromatogr B Biomed Sci Appl. 2001 May 5;755(1-2):37-46. doi: 10.1016/s0378-4347(00)00598-3.
3
Preparation and characterization of the F (ab)2 fragments of an aromatase activity-suppressing monoclonal antibody.
Steroids. 1997 Dec;62(12):776-81. doi: 10.1016/s0039-128x(97)00090-1.
4
On the interaction between single chain Fv antibodies and bacterial immunoglobulin-binding proteins.关于单链Fv抗体与细菌免疫球蛋白结合蛋白之间的相互作用
J Immunol Methods. 1994 Dec 28;177(1-2):151-63. doi: 10.1016/0022-1759(94)90152-x.
5
The interaction of proteins with hydroxyapatite. III. Mechanism.
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6
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Anal Biochem. 1984 Feb;136(2):433-9. doi: 10.1016/0003-2697(84)90240-9.
7
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