Purification process development for HER1 extracellular domain as a potential therapeutic vaccine.
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.
León
Dania
D
Process Development Department, Center for Molecular Immunology, 216th Street to 15th, P.O. Box 16040, Havana 11600, Cuba.
Prieto
Yadira
Y
Fernández
Eutimio G
EG
Pérez
Noemí
N
Montero
José A
JA
Palacios
Julio
J
Bulté
Dubhe
D
de la Luz
Kathya R
KR
Peña
Vladimir
V
Ferro
Williams
W
Sánchez
Belinda
B
Valdés
Rodolfo
R
Castillo
Adolfo
A
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
D000911:Antibodies, Monoclonal; D019496:Cancer Vaccines; C512478:EGFR protein, human; D066246:ErbB Receptors