Bioseparation最新文献_第7页

Expanded bed chromatography of proteins in small diameter columns. I. Scale down and validation. 小直径柱中蛋白质的扩展床层析。1 .缩小规模和验证。

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/a:1008193312969

S Ghose, H Chase

{"title":"Expanded bed chromatography of proteins in small diameter columns. I. Scale down and validation.","authors":"S Ghose, H Chase","doi":"10.1023/a:1008193312969","DOIUrl":"https://doi.org/10.1023/a:1008193312969","url":null,"abstract":"The use of large columns for expanded-bed chromatography in protein adsorption and purification can pose limitations in method scouting due to the high volumes of consumables involved in optimisation runs. Scaling down this technique would provide a practical and necessary first step for demonstrating its feasibility in very small beds. The performance of three columns of diameters 5.0, 1.0 and 0.5 cm were compared in terms of the bed expansion, hydrodynamics and breakthrough for lysozyme adsorption onto STREAMLINE-SP. This represented a scale-down factor of a 100-fold from the 5-cm column and the success was judged by the insignificant changes in performance based on the selected criteria. Bed characterisation and breakthrough runs indicated good plug flow behaviour, despite the high particle size to column diameter ratio in the smaller columns. The column efficiency was found to be sensitive to the vertical alignment, making it an important issue in scale down. The results of these investigations show that small diameter columns can be effectively used for mimicking the behaviour in scale up systems providing a useful tool for method scouting studies.","PeriodicalId":9179,"journal":{"name":"Bioseparation","volume":"9 1","pages":"21-8"},"PeriodicalIF":0.0,"publicationDate":"2000-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1008193312969","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21684693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 24

PCR identification of Salmonella cells in food and stool samples after immunomagnetic separation 食品和粪便中沙门氏菌免疫磁分离的PCR鉴定

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/A:1011125014586

A. Španová, B. Rittich, R. Karpíšková, L. Čechová, D. Škapová

引用次数: 52

Affinity chromatography of neoglycoproteins 新糖蛋白的亲和层析

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/A:1011187724356

Yucai Li, Eva Linné Larsson, H. Jungvid, I. Galaev, B. Mattiasson

引用次数: 20

Recovery of a monoclonal antibody from hybridoma culture supernatant by affinity precipitation with Eudragit S-100 用Eudragit S-100亲和沉淀法从杂交瘤培养上清中回收单克隆抗体

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/A:1011183904966

M. Ângela Taipa, R. Kaul, B. Mattiasson, J. Cabral

引用次数: 25

High-speed pectic enzyme fractionation by immobilised metal ion affinity membranes. 固定化金属离子亲和膜的高速果胶酶分离。

Bioseparation Pub Date : 2000-01-01 DOI: 10.1007/0-306-46889-1_22

S. Camperi, M. Grasselli, O. Cascone

引用次数: 4

Scale-up of recombinant cutinase recovery by whole broth extraction with PEG-phosphate aqueous two-phase 聚乙二醇-磷酸双水相全汤萃取法回收重组角质酶的研究

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/A:1008162209596

M. J. L. Costa, M. T. Cunha, J. Cabral, M. Aires-Barros

引用次数: 58

Aqueous polymer two-phase systems formed by new thermoseparating polymers. 新型热分离聚合物形成的水相聚合物体系。

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/a:1008167603733

J Persson, H O Johansson, I Galaev, B Mattiasson, F Tjerneld

{"title":"Aqueous polymer two-phase systems formed by new thermoseparating polymers.","authors":"J Persson, H O Johansson, I Galaev, B Mattiasson, F Tjerneld","doi":"10.1023/a:1008167603733","DOIUrl":"https://doi.org/10.1023/a:1008167603733","url":null,"abstract":"A set of new polymers that can be used as phase forming components in aqueous two-phase systems is presented. All polymers studied have thermoseparating properties i.e. form one separate polymer enriched phase and one aqueous solution when heated above the critical temperature. This property makes the polymers attractive alternatives to the polymers used in traditional aqueous two-phase systems such as poly(ethylene glycol) (PEG) and dextran. The thermal phase separation simplifies recycling of the polymers, thus making the aqueous two-phase systems more cost efficient and suitable for use in large scale. Thermoseparating polymers studied have been copolymers of ethylene oxide and propylene oxide (EO-PO), poly (N-isopropylacrylamide) (poly-NIPAM), poly vinyl caprolactam (poly-VCL) and copolymers of N-isopropylacrylamide and vinyl caprolactam with vinyl imidazole (poly(NIPAM-VI) and poly(VCL-VI), respectively). In addition, the copolymer poly(NIPAM-VI) has the property to be uncharged at pH above 7.0 and positively charged at lower pH. This allows the partitioning of protein to be directed by changing the pH in the system instead of the traditional addition of salt to direct the partitioning. Hydrophobically modified EO-PO copolymer (HM-(EO-PO)) with alkyl groups (C14) at both ends forms two-phase system with for example poly(NIPAM-VI). The phase diagram for poly(NIPAM-VI)/HM-(EO-PO) was determined and the model proteins lysozyme and BSA were partitioned in this system. For BSA in poly(NIPAM-VI)/HM-(EO-PO) system a change in pH from 8.0 to 5.4 results in a change of partition coefficient from K = 0.8 to K = 5.1, i.e. BSA could be transferred from the HM-(EO-PO) phase to the poly(NIPAM-VI) phase. BSA partitioning in poly(NIPAM-VI)/HM-(EO-PO) system allows quantitative BSA recovery, and recoveries of poly(NIPAM-VI) and HM-(EO-PO) were 53% and 92%, respectively, after the thermoseparation step.","PeriodicalId":9179,"journal":{"name":"Bioseparation","volume":"9 2","pages":"105-16"},"PeriodicalIF":0.0,"publicationDate":"2000-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1008167603733","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21734414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 59

Anion exchange purification of plasmid DNA using expanded bed adsorption. 扩展床吸附法纯化质粒DNA的阴离子交换。

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/a:1008134822673

G N Ferreira, J M Cabral, D M Prazeres

{"title":"Anion exchange purification of plasmid DNA using expanded bed adsorption.","authors":"G N Ferreira, J M Cabral, D M Prazeres","doi":"10.1023/a:1008134822673","DOIUrl":"https://doi.org/10.1023/a:1008134822673","url":null,"abstract":"Recent developments in gene therapy with non-viral vectors and DNA vaccination have increased the demand for large amounts of pharmaceutical-grade plasmid DNA. The high viscosity of process streams is of major concern in the purification of plasmids, since it can cause high back pressures in column operations, thus limiting the throughput. In order to avoid these high back pressures, expanded bed anion exchange chromatography was evaluated as an alternative to fixed bed chromatography. A Streamline 25 column filled with 100 ml of Streamline QXL media, was equilibrated with 0.5 M NaCl in TE (10 mM Tris, 1 mM EDTA, pH = 8.0) buffer at an upward flow of 300 cmh-1, E. coli lysates (obtained from up to 3 liters of fermentation broth) were injected in the column. After washing out the unbound material, the media was allowed to sediment and the plasmid was eluted with 1 M NaCl in TE buffer at a downward flow of 120 cmh-1. Purification factors of 36 +/- 1 fold, 26 +/- 0.4 plasmid purity, and close to 100% yields were obtained when less than one settled column volume of plasmid feed was injected. However, both recovery yield and purity abruptly decreased when larger amounts were processed-values of 35 +/- 2 and 5 +/- 0.7 were obtained for the recovery yield and purity, respectively, when 250 ml of feedstock were processed. In these cases, gel clogging and expansion collapse were observed. The processing of larger volumes, thus larger plasmid quantities, was only possible by performing an isopropanol precipitation step prior to the chromatographic step. This step led to an enhancement of the purification step.","PeriodicalId":9179,"journal":{"name":"Bioseparation","volume":"9 1","pages":"1-6"},"PeriodicalIF":0.0,"publicationDate":"2000-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1008134822673","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21684691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 51

The European Biotechnology Directory '99 Edited by Anita Crafts-Lightly and Ruth Willians 欧洲生物技术目录'99编辑安妮塔crafts - light和露丝威廉姆斯

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/A:1008142309063

M. Garaita, J. Kennedy

引用次数: 0

Handbook of Fourier Transform Raman and Infrared Spectra of Polymers A.H. Kuptsov and G.N. Zhizhin 高聚物的傅里叶变换拉曼和红外光谱手册

Bioseparation Pub Date : 2000-01-01 DOI: 10.1023/A:1008198824934

C. J. Knill

引用次数: 5