A Novel Method for Removing Polyethyleneimine from Biopharmaceutical Samples: Improving Assay Sensitivity of Residual DNA Qpcr | Zendy

Shumin Zhang | Zendy; Matthew Roberts | Zendy; Marisa Jones | Zendy; Jacob Zeitler | Zendy; Greg W. Kilby | Zendy; Aston Liu | Zendy; John R. White | Zendy

Open Access

A Novel Method for Removing Polyethyleneimine from Biopharmaceutical Samples: Improving Assay Sensitivity of Residual DNA Qpcr

Author(s) -

Shumin Zhang,

Matthew Roberts,

Marisa Jones,

Jacob Zeitler,

Greg W. Kilby,

Aston Liu,

John R. White

Publication year - 2020

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2020-0011

Subject(s) - dna , chromatography , genomic dna , biopharmaceutical , serial dilution , assay sensitivity , residual , microbiology and biotechnology , sodium dodecyl sulfate , alkaline lysis , polymerase chain reaction , chemistry , biology , plasmid , gene , biochemistry , medicine , dna vaccination , alternative medicine , pathology , algorithm , computer science

Polyethyleneimine (PEI) is a flocculent that is widely used in the downstream purification of monoclonal antibodies. It is an in-process residual that is carried through the drug purification process and strongly inhibits residual DNA quantitation by real-time quantitative PCR assay. Very high sample dilutions (e.g., 1:10,000) can overcome the interference of PEI, but at the cost of DNA assay sensitivity. Diluting samples poses a significant risk to the assay sensitivity needed to satisfy regulatory requirements on the quantitation of residual genomic DNA present per dose (i.e., 10 ng/dose). Removing PEI while retaining DNA, by the use of sodium dodecyl sulfate, heparin and/or sarkosyl can overcome the interference of PEI and allow a more accurate quantitation of residual DNA.

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