Development of a quantitative assay for residual host cell proteins in a recombinant subunit vaccine against human respiratory syncytial virus.
We have developed and validated a process-specific immunoligand assay based on the Threshold system for the quantification of residual host cell proteins (HCPs) in a recombinant subunit vaccine candidate against the human respiratory syncytial virus (hRSV). The industrial process of this vaccine produced in Escherichia coli, involved five chromatography steps for the production of clinical-grade batches. The clearance of non-product-related protein throughout the purification process was documented by the evaluation of the HCP content in the chromatographic fractions at each step of the downstream processing. The assay had a detection limit of 0.5 ng/ml of HCP equivalent to 10 parts per million (ppm). The quantification limit was 1.3 ng/ml of HCP, giving a sensitivity range of the assay of 10 to 30 ppm. To our knowledge, this is the first sensitive HCP assay reported for a vaccine.
Dagouassat
N
N
Centre d'Immunologie Pierre Fabre (CIPF), 5 Avenue Napoléon III, B.P. 497, F-74164, Saint-Julien-en-Genevois, France. nathalie.dagouassat@pierre-fabre.com
Haeuw
J F
JF
Robillard
V
V
Damien
F
F
Libon
C
C
Corvaïa
N
N
Lawny
F
F
Nguyen
T N
TN
Bonnefoy
J Y
JY
Beck
A
A
D000907:Antibodies, Bacterial; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000032:analysis / Q000276:immunology; D004340:Drug Contamination; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D066298:In Vitro Techniques; D012136:Respiratory Syncytial Viruses / Q000235:genetics* / Q000276:immunology*; D012680:Sensitivity and Specificity; D022223:Vaccines, Subunit / Q000032:analysis; D014614:Vaccines, Synthetic / Q000032:analysis; D014765:Viral Vaccines / Q000032:analysis*
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D014765:Viral Vaccines