Reports of Biochemistry and Molecular Biology

en Recombinant Protein Purification using Composite Polyacrylamide-Nanocrystalline Cryogel Monolith Column and Carbohydrate- Binding Module Family 64 as Affinity Tag بیوشیمی Biochemistry مقالات اصلی Original Article <div style="text-align: justify;">Background: In the field of recombinant protein production, downstream processing, especially protein purification, is critical and often the most expensive step. Carbohydrate binding module 64 (CBM64) was shown in 2011 to bind efficiently to a broad range of cellulose materials. Methods: In this study, we developed a protein purification method using nanocrystalline cellulose embedded in a polyacrylamide monolith cryogel and CBM64 affinity tag linked by intein to PD1 as a model protein. The CBM64-Intein-PD1 gene cassette was expressed in E. coli. Following cell lysis, CBM64-Intein-PD1 protein bound to the monolith PA-NCC cryogel. After washing and reducing the pH from 8.0 to 6.5, the intein underwent self-cleavage, resulting in the release and elution of pure PD1 protein. Results: The synthesized monolith column had a porous structure with an average pore size of 30 μm and a maximum binding capacity of 497 μg per gram of dried column. The yield of this purification method was 84%, while the yield of the His tag-acquired CBM64-Intein-PD1 method was 89%. Conclusions: We used cellulose as support for affinity chromatography, which can be used as a cost-effective method for protein purification.</div> Affinity tag, CBM64, PD1, Protein purification. 252 261 http://rbmb.net/browse.php?a_code=A-10-915-1&slc_lang=en&sid=1 Mohsen Danaeifar 100319475328460012201 100319475328460012201 No Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. Ziba Veisi Malekshahi 100319475328460012202 100319475328460012202 No Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. Fatemeh Kazemi-Lomedasht 100319475328460012203 100319475328460012203 No Department of Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. Mohammad Ali Mazlomi m-mazlomi@tums.ac.ir. 100319475328460012204 100319475328460012204 Yes Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.