+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Continuous desalting of refolded protein solution improves capturing in ion exchange chromatography: A seamless process

Continuous desalting of refolded protein solution improves capturing in ion exchange chromatography: A seamless process

Biotechnology Journal 12(6):

Truly continuous biomanufacturing processes enable an uninterrupted feed stream throughout the whole production without the need for holding tanks. We have utilized microporous anion and cation exchangers into which only salts, but not proteins, can penetrate into the pores for desalting of protein solutions, while diafiltration or dilution is usually employed for feed adjustments. Anion exchange and cation exchange chromatography columns were connected in series to remove both anions and cations. To increase operation performance, a continuous process was developed comprised of four columns. Continuous mode was achieved by staggered cycle operation, where one set of columns, consisting of one anion exchange and one cation exchange column, was loaded during the regeneration of the second set. Refolding, desalting and subsequent ion exchange capturing with a scFv as the model protein was demonstrated. The refolding solution was successfully desalted resulting in a consistent conductivity below 0.5 mS/cm from initial values of 10 to 11 mS/cm. With continuous operation process time could be reduced by 39% while productivity was increased to 163% compared to batch operation. Desalting of the protein solution resulted in up to 7-fold higher binding capacities in the subsequent ion exchange capture step with conventional protein binding resins.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 059546769

Download citation: RISBibTeXText

PMID: 28398709

DOI: 10.1002/biot.201700082

Related references

An integrated precipitation and ion-exchange chromatography process for antibody manufacturing: Process development strategy and continuous chromatography exploration. Journal of Chromatography. a 1533: 66-76, 2018

Method of calculation for the continuous mass exchange process between an antibiotic solution and ion-exchange resin. Pharmaceutical Chemistry Journal 12(1): 119-123, 1978

Separation of correctly refolded and mis-refolded consensus interferon by hydrophobic interaction chromatography. Sheng Wu Gong Cheng Xue Bao 21(3): 451-455, 2005

Characterization of MHC/peptide complexes refolded by a one-step ion-exchange chromatography. Journal of Immunological Methods 369(1-2): 81-90, 2011

Continuous process for the ion-exchange extraction of 6-aminopenicillanic acid from solution. Pharmaceutical Chemistry Journal 9(4): 247-251, 1975

Continuous process for the ion exchange extraction of 6 amino penicillanic acid from solution. Pharmaceutical Chemistry Journal (English Translation of Khimiko-Farmatsevticheskii Zhurnal) 9(4): (1976), 1975

Continuous purification of green fluorescent protein by simultaneous ion-exchange chromatography and size-exclusion chromatography. Abstracts of Papers American Chemical Society 221(1-2): BIOT 141, 2001

Sugar refinery liquor desalting by a new ion exchange process. International sugar journal 87(1039): 127-131, 1985

Desalting and buffer exchange of proteins using size-exclusion chromatography. Csh Protocols 2006(1):, 2006

Chromatography and desalting of nucleotides with ammonium bicarbonate on anion-exchange columns. Biochimica et Biophysica Acta 48(3): 0-590, 1961

Preparative desalting of bovine serum albumin by continuous annular chromatography. Journal of Chromatography A 763(1-2): 49-56, 1997

Recovery of potassium clavulanate from fermentation broth by ion exchange chromatography and desalting electrodialysis. Biotechnology and Bioprocess Engineering 14(6): 803-810, 2009

Improvement of crude oil desalting process in refinery electric desalting units. Chemistry and Technology of Fuels and Oils 19(1): 3-13, 1983

On-Line Ion Exchange Liquid Chromatography as a Process Analytical Technology for Monoclonal Antibody Characterization in Continuous Bioprocessing. Analytical Chemistry 89(21): 11357-11365, 2017

Preparation of casein phosphopeptides using a novel continuous process of combining an enzymatic membrane reactor with anion-exchange chromatography. Journal of Food Engineering 117(1): 105-112, 2013