Your browser doesn't support javascript.
loading
N terminal sequencing for practical detection of monoclonal antibody / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1473-1480, 2014.
Artigo em Chinês | WPRIM | ID: wpr-345577
ABSTRACT
Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Peptídeos / Espectrometria de Massas / Mapeamento de Peptídeos / Cromatografia por Troca Iônica / Sequência de Aminoácidos / Análise de Sequência de Proteína / Focalização Isoelétrica / Métodos / Anticorpos Monoclonais Tipo de estudo: Estudo diagnóstico / Estudo prognóstico Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Biotechnology Ano de publicação: 2014 Tipo de documento: Artigo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Peptídeos / Espectrometria de Massas / Mapeamento de Peptídeos / Cromatografia por Troca Iônica / Sequência de Aminoácidos / Análise de Sequência de Proteína / Focalização Isoelétrica / Métodos / Anticorpos Monoclonais Tipo de estudo: Estudo diagnóstico / Estudo prognóstico Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Biotechnology Ano de publicação: 2014 Tipo de documento: Artigo