Your browser doesn't support javascript.
loading
Quantitation of the monoclonal antibody Denosumab by bioassay and validated LC methods.
Perobelli, Rafaela Ferreira; Xavier, Bruna; Silveira, Alice Rosa da; Remuzzi, Gabriel Lunardi; Motta, Luís Gustavo Jung; Dalmora, Sérgio Luiz.
Afiliação
  • Perobelli RF; Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
  • Xavier B; Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
  • Silveira ARD; Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
  • Remuzzi GL; Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
  • Motta LGJ; Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
  • Dalmora SL; Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil. Electronic address: sdalmora@terra.com.br.
Int J Biol Macromol ; 119: 96-104, 2018 Nov.
Article em En | MEDLINE | ID: mdl-30048724
The monoclonal antibody Denosumab (DmAb) is clinically used to treat osteoporosis and bone loss. We developed a bioassay based on the ability of DmAb to inhibit the effect of human receptor activator of nuclear factor-κB ligand (RANKL) to stimulate the formation of osteoclasts derived from RAW 264.7 cells. This bioassay was applied in conjunction with size exclusion high-performance liquid chromatography (SE-HPLC) and reversed-phase high-performance liquid chromatography (RP-HPLC) methods, with diode array detection (DAD), validated for the quantitation of this biotechnology-derived medicine. The SE-HPLC(DAD) method was carried out on a TSKGel G2000SWXL column and the mobile phase consisted of potassium phosphate buffer with sodium chloride, pH 7.4. The gradient RP-HPLC(DAD) method was carried out on a Vydac 214TP C4 column at 60 °C. The mobile phases consisted of 0.1% v/v trifluoroacetic acid (TFA) in water and 0.1% v/v TFA in acetonitrile. Calibration curves were linear over the concentration ranges 6-200 µg mL-1 and 6-300 µg mL-1 for the SE-HPLC(DAD) and RP-HPLC(DAD) methods respectively. The bioassay results correlated with the LC methods results, indicating the capabilities of these methods to quantitate DmAb, which will contribute to ensure the batch-to-batch consistency and efficacy of this biotherapeutic.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bioensaio / Cromatografia Líquida de Alta Pressão / Denosumab / Anticorpos Monoclonais Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bioensaio / Cromatografia Líquida de Alta Pressão / Denosumab / Anticorpos Monoclonais Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda