RESUMO

HER1 is a tumor associated antigen emerging as an attractive target for cancer therapy. In the present study we demonstrated for first time that HER1 extracellular domain can be purified by a downstream process at pilot scale based on immunoaffinity chromatography from bioreactor supernatant of HEK 293 transfectomes. Filtered supernatant was applied to CNBr-activated Sepharose CL-4B with monoclonal antibody anti-human EGF immobilized, followed by three additional chromatographic polishing steps. HER1 extracellular domain was obtained with high purity (>95%), low DNA content, and biological activity.

Assuntos

Cromatografia de Afinidade/métodos , Receptores ErbB/isolamento & purificação , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Vacinas Anticâncer/química , Vacinas Anticâncer/imunologia , Linhagem Celular , Cromatografia em Gel , Cromatografia por Troca Iônica , Receptores ErbB/química , Receptores ErbB/genética , Receptores ErbB/imunologia , Citometria de Fluxo , Humanos , Estrutura Terciária de Proteína , Espectrometria de Massas em Tandem , Transfecção