Abstract
Methods suitable for purification of IgM monoclonal antibodies from serum-free tissue culture supernatants are described in this case study. We review techniques found to be useful in purifying proteins and the techniques applied to establish utility. Partitioning techniques include polyethylene glycol (PEG) precipitation, size exclusion chromatography, anion and cation exchange chromatography, hydroxylapatite chromatography, and immunoaffinity. Modification techniques include the use of enzymes (e.g. DNAse). Protein purity is achieved primarily with precipitation, size exclusion chromatography, and immunoaffinity. DNA removal is greatest with anion exchange, immunoaffinity,and a combination of DNAse and size exclusion chromatography. Virus is partitioned most effectively through hydroxylapatite and immunoaffinity. A cascade of several appropriate steps provides contaminant protein clearance of >100x (purity greater than 99%), DNA clearance of >1,000,000x, and virus clearance of >100,000x.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
