chromatography-2017-posters-accepted-abstracts

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Volume 8, Issue 5 (Suppl)

J Chromatogr Sep Tech, an open access journal

ISSN: 2157-7064

Chromatography 2017

August 07-09, 2017

August 07-09, 2017 | Rome, Italy

World Congress on

Chromatography

J Chromatogr Sep Tech 2017, 8:5(Suppl)

DOI: 10.4172/2157-7064-C1-032

Novel technology for protein capture: Mixed-mode expanded-bed adsorption

Dong-Qiang Lin

and

Shan-Jing Yao

Zhejiang University, China

or biopharmaceuticals downstream processes, it always requires highly productive and robust technologies to improve the process

efficiency. Expanded-bed adsorption (EBA) is an innovative technology that allows capturing proteins directly from un-clarified

feedstock, such as cell culture broth and homogenization. EBA technology combines solid-liquid separation with an adsorption step

in a single-unit operation, aiming at increased overall yield, reduced operational time, and less requirements for capital investment

and consumables. Mixed-mode chromatography (MMC) is a novel technology for bio-product separation, which combines multiple

binding modes like hydrophobic and electrostatic interactions, hydrogen bonding, etc. High capacity, salt-tolerance, good selectivity

and relatively low cost are the major advantages of MMC for direct capture process. In the present work two chromatographic

techniques, EBA and MMC, were integrated to develop new separation technology, mixed-mode EBA, improving the protein capture

efficiency and reducing the pretreatments on the feedstock, such as clarification, dilution and salt-adjustment. Several MMC ligands

were coupled onto typical matrices (densified agarose beads) for EBA. The static adsorption, adsorption kinetics and dynamic

binding were investigated, and the effects of pH and salt addition were evaluated. New technology was challenged with two typical

biopharmaceutical processes, monoclonal antibody (mAb) capture from CHO cell culture broth and recombinant human albumin

serum (rHSA) isolation from Pichia pastoris fermentation broth. After the optimization of operation conditions, high separation

efficiency (purity, recovery, productivity) was obtained. The results demonstrated that mixed-mode EBA, combining the advantages

of EBA and MMC, would be a promising new platform for protein capture with reduced feedstock pretreatments, high efficiency and

relative low cost. New technology developed in the present work could also be expanded to other bio-product processes.

lindq@zju.edu.cn

Adsorptive removal of Triton X-100 from human plasma and its derivatives

Gargi Redkar

and

Sandeep Kale

DBT-ICT-Centre for Energy Biosciences, India

Institute of Chemical Technology, Maharashtra, India

iral transmission during the use of human plasma and its derivatives to treat various medical conditions can be fatal. Solvent/

detergent treatment using non-ionic detergents like Triton X-100 inactivate the lipid enveloped viruses. However, the detergent

interferes with downstream processing and analysis. Also, WHO permits a residual level of <25 ppm of Triton X-100 and thus it needs

to be removed from post viral inactivation. Removal of Triton X-100 poses a challenge due to its low CMC and non-ionic character.

Selective removal of Triton X-100 was studied using various hydrophobic resins screened on the basis of adsorption capacity, uptake

kinetics and effect of plasma proteins on these parameters. Resins showing higher adsorption capacity and uptake rate with lower

protein binding were selected for column studies. Breakthrough capacity of the shortlisted resins was determined at different flow

rates and concentrations along with the effect of proteins. A simple and sensitive HPLC method was developed to detect Triton X-100

in the treated samples at ppm level. This research work asserted the impact of various resin characteristics and plasma proteins on

selective detergent removal and thus the mechanism of adsorption of Triton X-100 onto these resins.

sb.kale@ictmumbai.edu.in