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Combined polyethylene glycol and CaCl2 precipitation for the capture and purification of recombinant antibodies

Paper ID Volume ID Publish Year Pages File Format Full-Text
34461 45026 2009 9 PDF Available
Title
Combined polyethylene glycol and CaCl2 precipitation for the capture and purification of recombinant antibodies
Abstract

•Direct capture step for the purification of recombinant antibodies from clarified culture supernatants.•Combination of CaCl2 and polyethylene glycol (PEG) precipitation equivalent to protein A affinity chromatography.•Basis for a platform process to capture antibodies from a clarified culture supernatant.

A direct capture step was developed for the purification of recombinant antibodies from clarified culture supernatants. This method is a combination of CaCl2 and polyethylene glycol (PEG) precipitation. For separation of high molecular weight impurities, such as dsDNA and aggregates, CaCl2 precipitation was the method of choice, and PEG precipitation was used for monoclonal antibody (mAb) capturing to separate low molecular weight impurities, mainly host cell proteins (HCP). Precipitation tests were performed with five different CHO cell culture supernatants, and showed yields of at least 80–95%. Purity was determined by size exclusion chromatography, HCP ELISA, and two-dimensional differential in-gel electrophoresis (2D DIGE); the measured values from CaCl2/PEG precipitation were comparable to those from Protein A purification. These results suggest that this novel purification method could be used as the basis for a platform process to capture antibodies from a clarified culture supernatant. CaCl2/PEG precipitation provides enough selectivity and recovery to substantiate replacing the Staphylococcus Protein A affinity chromatography step of the antibody purification process.

Keywords
Protein A; mAb; PEG; Polyethylene glycol; Precipitation; CHO
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Combined polyethylene glycol and CaCl2 precipitation for the capture and purification of recombinant antibodies
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 49, Issue 11, November 2014, Pages 2001–2009
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us