Process characterization for metal-affinity chromatography of an Fc fusion protein: a design-of-experiments approach

2001 ◽  
Vol 34 (2) ◽  
pp. 71 ◽  
Author(s):  
Abhinav A. Shukla ◽  
Laura Sorge ◽  
Joshua Boldman ◽  
Steve Waugh
Keyword(s):  
Fusion Protein ◽  
Design Of Experiments ◽  
Affinity Chromatography ◽  
Protein A ◽  
Metal Affinity ◽  
Process Characterization ◽  
Fc Fusion Protein

Quality by Design Applied to a Fc-Fusion Protein: A Case Study

2013 ◽  
pp. 155-189 ◽  
Author(s):  
Alex Eon-Duval ◽  
Ralf Gleixner ◽  
Pascal Valax ◽  
Miroslav Soos ◽  
Benjamin Neunstoecklin ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Quality By Design ◽  
Protein A ◽  
Fc Fusion Protein

scholarly journals Favorable pharmacokinetics in hemophilia B for nonacog beta pegol versus recombinant factor IX-Fc fusion protein: A randomized trial

2019 ◽  
Vol 3 (2) ◽  
pp. 268-276 ◽  
Author(s):  
Carmen Escuriola Ettingshausen ◽  
Inga Hegemann ◽  
Mindy L. Simpson ◽  
Adam Cuker ◽  
Roshni Kulkarni ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Randomized Trial ◽  
Protein A ◽  
Factor Ix ◽  
Hemophilia B ◽  
Fc Fusion Protein ◽  
Recombinant Factor Ix

scholarly journals Positive Selection of Specific Antibodies Produced against Fusion Proteins

2020 ◽  
Vol 3 (2) ◽  
pp. 37
Author(s):  
Lukas Kramberger-Kaplan ◽  
Tina Austerlitz ◽  
Holger Bohlmann
Keyword(s):  
Fusion Protein ◽  
Affinity Chromatography ◽  
Positive Selection ◽  
Fusion Proteins ◽  
Protein A ◽  
High Specificity ◽  
Western Blots ◽  
Specific Antibodies ◽  
E Coli ◽  
Selection Of

A method for the positive selection of specific antibodies for target proteins expressed as fusion proteins for the production of antiserum is presented. As proof of concept, the fusion protein FLAG::His::GFP::His::FLAG was expressed in Escherichia coli, purified, and used for the immunization of rabbits. The obtained serum was precleared via protein A affinity. A CusF::FLAG fusion protein was expressed in the periplasm of E. coli and purified. GFP without tags was also expressed in E. coli and purified via organic extraction. These proteins were then coupled to NHS-activated sepharose and used for the positive selection of Anti-GFP and Anti-FLAG antibodies. The obtained sera were tested for their specificity against different protein samples and fusion proteins in Western blots. A high specificity of the antibodies could be achieved by a single affinity chromatography step. In general, we advise to express the target protein with different tags and in different E. coli compartments for antibody production and affinity chromatography.

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