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Determination of free sulfhydryl contents for proteins including monoclonal antibodies by use of SoloVPE.
Zhang, Yuling; Qi, Pei.
Afiliação
  • Zhang Y; Analytical and Formulation Department, Lundbeck Seattle BioPharmaceuticals, Inc, 11804 North Creek Parkway South, Bothell, Washington, 98011, United States. Electronic address: yuzn@lundbeck.com.
  • Qi P; Analytical and Formulation Department, Lundbeck Seattle BioPharmaceuticals, Inc, 11804 North Creek Parkway South, Bothell, Washington, 98011, United States.
J Pharm Biomed Anal ; 201: 114092, 2021 Jul 15.
Article em En | MEDLINE | ID: mdl-33984827
Free sulfhydryls are important properties of protein products including monoclonal antibodies (mAbs). Here, a new technology, variable pathlength extension (SoloVPE), is employed to quantify the amount of free sulfhydryl in monoclonal antibodies (mAbs) using the well-known Ellman reagent. Briefly, the unbound thiols (free sulfhydryls) of proteins including mAbs react with Ellman reagent to produce a 2-nitro-5-thiobenzoate (TNB2-) which is detected at visible wavelength of 412 nm and quantified. The method does not require dilution of antibody samples, is simple, reproducible and takes less than one hour to complete. Values obtained by the new method are compared to literature values from traditional UV or fluorescence methods with agreements. Qualification and trending data over two years of method utilization in our labs support that assay variability is minimal with an intermediate precision of relative standard deviation (RSD) ≤ 10 % and a limit of quantification (LOQ) of 0.1 mol/mol, which is sufficient to measure free sulfhydryl content in proteins including mAbs.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Antineoplásicos Imunológicos / Anticorpos Monoclonais Idioma: En Revista: J Pharm Biomed Anal Ano de publicação: 2021 Tipo de documento: Article País de publicação: Reino Unido

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Antineoplásicos Imunológicos / Anticorpos Monoclonais Idioma: En Revista: J Pharm Biomed Anal Ano de publicação: 2021 Tipo de documento: Article País de publicação: Reino Unido