为了方便更多的研究人员系统的学习和了解蛋白纯化技术，生物库会在随后的时间内陆续的推出GE公司蛋白纯化相关技术手册免费提供下载，由于中文手册文件较大，如需中文版技术手册请注册本站评论回复告知邮箱，或点击关注本站新浪微博后发私信给生物库告知您的邮箱，生物库会第一时间Email给您！同时也感谢您对本站的支持与关注！

GE技术手册发布计划：

第一期：
Ion Exchange Chromatography & Chromatofocusing Principles and Methods
离子交换色谱及色谱聚焦原理和方法

第二期：
Hydrophobic Interaction and Reversed Phase Chromatography Principles and Methods
疏水相互作用和反向层析技术原理和方法

第三期：
Gel Filtration Principles and Methods
凝胶过滤原理和方法

第四期：
Affinity Chromatography Principles and Methods
亲和色谱原理和方法

第五期：
Antibody Purification Handbook
抗体纯化手册

第六期：
Cell Separation Media Methodology and applications
细胞分离介质方法学和应用

第七期：
Expanded bed adsorption Principles and Methods

第八期：
Protein Purification Handbook
蛋白质纯化手册

第九期：
Microcarrier Cell Culture Principles and Methods

第十期：
Recombinant Protein Purification Handbook Principles and Methods
重组蛋白纯化手册原理和方法

第十一期:
Isolation of mononuclear cells Methodology and applications
单核细胞的分离方法与应用

第十二期：
GST Gene Fusion System Handbook

第十三期：
Purifying Challenging Proteins Principles and Methods
富有挑战性的蛋白质纯化原理和方法

第十四期：
2-D Electrophoresis Principles and Methods
2-D双向电泳的原理与方法

今天我们就发布第五期：
Antibody Purification Handbook
抗体纯化手册

英文版：Antibody Purification Handbook

Contents
Introduction............................................................................................................................................................... 5
Chapter 1. Antibody structure, classification, and production............................... 7
Native sources......................................................................................................................................................... 7
Genetically engineered sources.....................................................................................................................11
References...............................................................................................................................................................13
Chapter 2. Sample preparation.........................................................................................................15
Sources and their associated contaminants...........................................................................................15
Extraction of recombinant antibodies and antibody fragments....................................................16
Clarification of serum, ascites, cell culture supernatant, or cell lysate.......................................18
Sample preparation before purification.....................................................................................................19
Desalting and buffer exchange......................................................................................................................24
Chapter 3. Small-scale purification by affinity chromatography.......................43
General considerations.....................................................................................................................................43
Adding a polishing step after intitial purification...................................................................................45
Protein G and protein A bind to different IgG...........................................................................................47
Purification using Protein G Sepharose media........................................................................................49
Purifying Fab and F(ab’)2 fragments.............................................................................................................53
Multiwell plates and columns prepacked with Protein G Sepharose High Performance....54
Purification using protein A-based media.................................................................................................66
MabSelect media and prepacked columns..............................................................................................84
Purification of other classes of antibodies................................................................................................95
Making immunospecific purification media with custom ligands..............................................100
Chapter 4. Removal of specific contaminants after initial purification............105
Bovine immunoglobulins...............................................................................................................................105
Albumin and transferrin ................................................................................................................................106
a2-macroglobulin and haptoglobulin ......................................................................................................109
Dimers and aggregates..................................................................................................................................109
DNA and endotoxins........................................................................................................................................110
Affinity ligands....................................................................................................................................................111
Chapter 5. Automated purification of antibodies using
ÄKTAdesign systems...............................................................................................................................113
Chapter 6. Multistep purification strategies......................................................................117
Examples of multistep purification............................................................................................................117
Handbook 18-1037-46 AD
Chapter 7. Large-scale purification ..........................................................................................125
Platform technologies in MAb purification.............................................................................................125
Process design....................................................................................................................................................126
High productivity media for MAb purification.......................................................................................127
Prepacked, disposable solutions speed up the downstream process......................................127
Custom Designed Media and columns....................................................................................................128
Appendix 1. Characteristics of Protein G and Protein A Sepharose
products.............................................................................................................................................................129
Appendix 2. Analytical assays during purification........................................................137
Appendix 3. Immunoprecipitation techniques..................................................................139
Appendix 4. General instructions for affinity purification
using HiTrap columns............................................................................................................................142
Appendix 5. Column packing and preparation ................................................................144
Appendix 6. Storage of biological samples..........................................................................147
Appendix 7. Converting from linear flow (cm/h) to volumetric
flow rates (ml/min) and vice versa..............................................................................................148
Appendix 8. Conversion data: proteins, column pressures....................................149
Appendix 9. Principles and standard conditions for
different purification techniques..................................................................................................150
Product index................................................................................................................................................157
Related literature......................................................................................................................................158
Ordering information.............................................................................................................................159

更多内容请点击下载：Antibody Purification Handbook

中文版：抗体纯化手册

如需中文版技术手册请注册本站评论回复告知邮箱，或点击关注本站新浪微博后发私信给我告知您的邮箱，我会第一时间Email给您！同时也感谢您对本站的支持与关注！