alexa The Comparison of Chemiluminescent- and Colorimetric-detection Based ELISA for Chinese Hamster Ovary Host Cell Proteins Quantification in Biotherapeutics
ISSN: 2155-9821

Journal of Bioprocessing & Biotechniques
Open Access

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Research Article

The Comparison of Chemiluminescent- and Colorimetric-detection Based ELISA for Chinese Hamster Ovary Host Cell Proteins Quantification in Biotherapeutics

Fengqiang Wang*, Dennis Driscoll, Daisy Richardson and Alexandre Ambrogelly
Bioprocess Development, Merck Research Laboratories, Union, NJ, USA
Corresponding Author : Fengqiang Wang, PhD
Associate Principal Scientist
Bioprocess Development
Merck Research Laboratories, NJ, USA
Tel: 908-740-5106
Fax: 908-740-4366
Email: [email protected]
Received September 12, 2013; Accepted October 21, 2013; Published October 30, 2013
Citation: Wang F, Driscoll D, Richardson D, Ambrogelly A (2013) The Comparison of Chemiluminescent- and Colorimetric-detection Based ELISA for Chinese Hamster Ovary Host Cell Proteins Quantification in Biotherapeutics. J Bioprocess Biotech 3:136 doi: 10.4172/2155-9821.1000136
Copyright: © 2013 Wang F, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Biologics manufacturing requires the clearance of Host Cell Proteins (HCPs) from recombinant therapeutic protein to acceptable low levels to ensure product purity and patient safety. To ensure adequate removal, a highly sensitive method, commonly in the form of Enzyme-Linked Immunosorbent Assay (ELISA), is necessary to quantify the HCPs amount in process intermediates and drug substance. We report the development of a chemiluminescent detection based ELISA (luminescent ELISA) in lieu of previously used colorimetric method (colorimetric ELISA) to improve assay sensitivity for the quantification of Chinese Hamster Ovary (CHO) HCPs in a monoclonal antibody product (mAb-A). For luminescent ELISA, Pierce Supersignal ELISA Femto was chosen as the substrate to replace colorimetric substrate TMB. The assay performance of luminescent and colorimetric ELISA was directly compared side-by-side. Our data show that luminescent ELISA has better signal/background ratio, broader linear range over logarithmic scales, and better linearity within the same linear range than colorimetric ELISA. Luminescent ELISA also demonstrates better lowend linearity, greater accuracy and precision. In addition, the Limit of Detection (LOD) and Limit of Quantification (LOQ) are significantly improved with luminescent ELISA as compared to colorimetric ELISA. In summary, luminescent ELISA is a more sensitive method and demonstrates superiority over colorimetric method for CHO HCP quantification.

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