Automated sample preparation for CE-SDS

被引:12
作者
Le, M. Eleanor [1 ]
Vizel, Alona [1 ]
Hutterer, Katariina M. [1 ]
机构
[1] Amgen Inc, Proc & Prod Dev, Thousand Oaks, CA 91320 USA
关键词
Automation; CE-SDS; Capillary gel electrophoresis; Phynexus; CAPILLARY-ELECTROPHORESIS; PURIFICATION; REDUCTION; PROTEIN; ONLINE;
D O I
10.1002/elps.201200644
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Traditionally, CE with SDS (CE-SDS) places many restrictions on sample composition. Requirements include low salt content, known initial sample concentration, and a narrow window of final sample concentration. As these restrictions require buffer exchange for many sample types, sample preparation is often tedious and yields poor sample recoveries. To improve capacity and streamline sample preparation, an automated robotic platform was developed using the PhyNexus Micro-Extractor Automated Instrument (MEA) for both the reduced and nonreduced CE-SDS assays. This automated sample preparation normalizes sample concentration, removes salts and other contaminants, and adds the required CE-SDS reagents, essentially eliminating manual steps during sample preparation. Fc-fusion proteins and monoclonal antibodies were used in this work to demonstrate benefits of this approach when compared to the manual method. With optimized conditions, this application has demonstrated decreased analyst hands on time and reduced total assay time. Sample recovery greater than 90% can be achieved, regardless of initial composition and concentration of analyte.
引用
收藏
页码:1369 / 1374
页数:6
相关论文
共 16 条
[1]  
Elisabeth R., 2006, PROCESS SCALE BIOSEP, P1
[2]   Technology trends in antibody purification [J].
Gagnon, Pete .
JOURNAL OF CHROMATOGRAPHY A, 2012, 1221 :57-70
[3]   Improved solid-phase microextraction device for use in on-line immunoaffinity capillary electrophoresis [J].
Guzman, NA .
ELECTROPHORESIS, 2003, 24 (21) :3718-3727
[4]   Targeted codon optimization improves translational fidelity for an Fc fusion protein [J].
Hutterer, Katariina M. ;
Zhang, Zhongqi ;
Michaels, Mark Leo ;
Belouski, Ed ;
Hong, Robert W. ;
Shah, Bhavana ;
Berge, Mark ;
Barkhordarian, Hedieh ;
Le, Eleanor ;
Smith, Steve ;
Winters, Dwight ;
Abroson, Frank ;
Hecht, Randy ;
Liu, Jennifer .
BIOTECHNOLOGY AND BIOENGINEERING, 2012, 109 (11) :2770-2777
[5]   SUPPORTED LIQUID MEMBRANE TECHNIQUES FOR SAMPLE PREPARATION AND ENRICHMENT IN ENVIRONMENTAL AND BIOLOGICAL ANALYSIS [J].
JONSSON, JA ;
MATHIASSON, L .
TRAC-TRENDS IN ANALYTICAL CHEMISTRY, 1992, 11 (03) :106-114
[6]   Hepcidin Revisited, Disulfide Connectivity, Dynamics, and Structure [J].
Jordan, John B. ;
Poppe, Leszek ;
Haniu, Mitsuru ;
Arvedson, Tara ;
Syed, Rashid ;
Li, Vivian ;
Kohno, Hiko ;
Kim, Helen ;
Schnier, Paul D. ;
Harvey, Timothy S. ;
Miranda, Les P. ;
Cheetham, Janet ;
Sasu, Barbra J. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2009, 284 (36) :24155-24167
[7]   Mechanism of Antibody Reduction in Cell Culture Production Processes [J].
Kao, Yung-Hsiang ;
Hewitt, Daniel P. ;
Trexler-Sehrnidt, Melody ;
Laird, Michael W. .
BIOTECHNOLOGY AND BIOENGINEERING, 2010, 107 (04) :622-632
[8]   Analysis of monoclonal antibody product heterogeneity resulting from alternate cleavage sites of signal peptide [J].
Kotia, Ruchi B. ;
Raghani, Anil R. .
ANALYTICAL BIOCHEMISTRY, 2010, 399 (02) :190-195
[9]   Automated sample preparation and analysis using a sequential-injection-capillary electrophoresis (SI-CE) interface [J].
Kulka, Stephan ;
Quintas, Guillermo ;
Lendl, Bernhard .
ANALYST, 2006, 131 (06) :739-744
[10]  
Li F., 1995, BIOPROCESS J, V4, P1