Development of a multiplexing method for the quantification of "high-risk" host cell lipases in biotherapeutics by Luminex.

Autor: Osko JD; Analytical Research & Development, Merck & Co., Inc., Rahway, NJ, USA. Electronic address: jeremy.osko@merck.com., Rivera S; Analytical Research & Development, Merck & Co., Inc., Rahway, NJ, USA., Wang F; Analytical Research & Development, Merck & Co., Inc., Rahway, NJ, USA., Niedringhaus T; Analytical Research & Development, Merck & Co., Inc., Rahway, NJ, USA.
Jazyk: angličtina
Zdroj: Analytica chimica acta [Anal Chim Acta] 2024 Dec 15; Vol. 1332, pp. 343349. Date of Electronic Publication: 2024 Oct 21.
DOI: 10.1016/j.aca.2024.343349
Abstrakt: Clearance of residual Host Cell Proteins (HCPs) is critical for the manufacturing processes of biotherapeutics. HCPs have the potential to impact product efficacy and quality, posing a risk to patient safety. It is therefore essential to be able to both identify and quantitate HCPs throughout drug development, even if the proteins are present in low concentrations. Traditional Enzyme-Linked Immunosorbent Assays (ELISAs) have historically served as the gold standard for monitoring HCPs; however, ELISA methods are labor-intensive and costly. With an increase of HCPs being identified below detectable quantification levels, there is a need for simultaneous detection of selectively targeted HCPs. Here, we develop a Luminex multiplexing method that is able to accurately quantify two "high-risk" lipases Lipoprotein Lipase (LPL) and Phospholipase B-Like 2 (PLBL2) within the same assay. This study outlines the method development for optimizing parameters such as antibody constructs, conjugation ratios, signal enhancement, and more in order to create the most efficient multiplexing method. As a result, a Luminex multiplexing method can provide a similar result to a monoplexing ELISA method but in a faster and more cost-effective manner. This method can be expanded to include other "high-risk" HCPs and used for future HCP applications.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
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