title: Purification process development for HER1 extracellular domain as a potential therapeutic vaccine.
abstract: 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.
authors.affiliation: Process Development Department, Center for Molecular Immunology, 216th Street to 15th, P.O. Box 16040, Havana 11600, Cuba.
authors.forename: Dania
authors.initials: D
authors.lastname: León
authors.forename: Yadira
authors.initials: Y
authors.lastname: Prieto
authors.forename: Eutimio G
authors.initials: EG
authors.lastname: Fernández
authors.forename: Noemí
authors.initials: N
authors.lastname: Pérez
authors.forename: José A
authors.initials: JA
authors.lastname: Montero
authors.forename: Julio
authors.initials: J
authors.lastname: Palacios
authors.forename: Dubhe
authors.initials: D
authors.lastname: Bulté
authors.forename: Kathya R
authors.initials: KR
authors.lastname: de la Luz
authors.forename: Vladimir
authors.initials: V
authors.lastname: Peña
authors.forename: Williams
authors.initials: W
authors.lastname: Ferro
authors.forename: Belinda
authors.initials: B
authors.lastname: Sánchez
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Valdés
authors.forename: Adolfo
authors.initials: A
authors.lastname: Castillo
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D019496:Cancer Vaccines / Q000737:chemistry / Q000276:immunology; D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D066246:ErbB Receptors / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D005434:Flow Cytometry; D006801:Humans; D017434:Protein Structure, Tertiary; D053719:Tandem Mass Spectrometry; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D019496:Cancer Vaccines; C512478:EGFR protein, human; D066246:ErbB Receptors