Synthetic polymers are more effective than natural flocculants for the clarification of tobacco leaf extracts

被引:11
作者
Buyel, Johannes F. [1 ]
Fischer, Rainer [1 ,2 ]
机构
[1] Rhein Westfal TH Aachen, Inst Mol Biotechnol, Worringerweg 1, D-52074 Aachen, Germany
[2] Fraunhofer Inst Mol Biol & Appl Ecol, D-52074 Aachen, Germany
基金
欧洲研究理事会;
关键词
Bioprocess development; Plant-derived biopharmaceuticals; Protein purification; Plant extract flocculation; Design of experiments (DoE); SOLID-LIQUID SEPARATION; OLIVE MILL; POLYELECTROLYTE FLOCCULATION; WASTE-WATER; PURIFICATION; COAGULATION; RECOVERY; ADSORPTION; EFFICACY; CHITOSAN;
D O I
10.1016/j.jbiotec.2014.12.018
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The use of synthetic polymers as flocculants can increase filter capacity and thus reduce the costs of downstream processing during the production of plant-derived biopharmaceutical proteins, but this may also attract regulatory scrutiny due to the potential toxicity of such compounds. Therefore, we investigated the efficacy of three non-toxic natural flocculants (chitosan, kaolin and polyphosphate) alone and in combination with each other or with a synthetic polymer (Polymin P) during the clarification of tobacco leaf extracts. We used a design-of-experiments approach to determine the impact of each combination on filter capacity. We found that Polymin P was most effective when used on its own but the natural flocculants were more effective when used in combination. The combination of chitosan and polyphosphate was the most effective natural flocculant, and this was identified as a potential replacement for Polymin P under neutral and acidic extraction conditions independent of the conductivity, even though the efficiency of flocculation was lower than for Polymin P. None of the tested flocculants reduced the concentration of total soluble protein in the feed stream or the recovery of the model fluorescent protein DsRed. (C) 2014 Elsevier B.V. All rights reserved.
引用
收藏
页码:37 / 42
页数:6
相关论文
共 39 条
[1]  
Anderson M.J., 2000, DOE SIMPLIFIED PRACT
[2]  
[Anonymous], 2006, CURR PROTOC MOL BIOL, DOI DOI 10.11.11-10.11A.19
[3]   Micro total analysis systems. 2. Analytical standard operations and applications [J].
Auroux, PA ;
Iossifidis, D ;
Reyes, DR ;
Manz, A .
ANALYTICAL CHEMISTRY, 2002, 74 (12) :2637-2652
[4]  
Benea M., 2004, STUD UBB GEOL, V49, P33
[5]   Generic chromatography-based purification strategies accelerate the development of downstream processes for biopharmaceutical proteins produced in plants [J].
Buyel, Johannes F. ;
Fischer, Rainer .
BIOTECHNOLOGY JOURNAL, 2014, 9 (04) :566-577
[6]   Scale- down models to optimize a filter train for the downstream purification of recombinant pharmaceutical proteins produced in tobacco leaves [J].
Buyel, Johannes F. ;
Fischer, Rainer .
BIOTECHNOLOGY JOURNAL, 2014, 9 (03) :415-425
[7]   Flocculation increases the efficacy of depth filtration during the downstream processing of recombinant pharmaceutical proteins produced in tobacco [J].
Buyel, Johannes F. ;
Fischer, Rainer .
PLANT BIOTECHNOLOGY JOURNAL, 2014, 12 (02) :240-252
[8]   Downstream processing of biopharmaceutical proteins produced in plants The pros and cons of flocculants [J].
Buyel, Johannes Felix ;
Fischer, Rainer .
BIOENGINEERED, 2014, 5 (02) :138-142
[9]   Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study [J].
Buyel, Johannes Felix ;
Fischer, Rainer .
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2014, (83)
[10]   Polyphosphates as inorganic polyelectrolytes interacting with oppositely charged ions, polymers and deposited on surfaces: fundamentals and applications [J].
Cini, N. ;
Ball, V. .
ADVANCES IN COLLOID AND INTERFACE SCIENCE, 2014, 209 :84-97