caa a22 4500 477127169 CHVBK 20180405111653.0 cr unu---uuuuu 170330e19971001xx s 000 0 eng 10.1023/A:1012168621337 doi (NATIONALLICENCE)springer-10.1023/A:1012168621337 Development and Use of Enzyme-Linked Immunosorbent Assays (ELISA) for the Detection of Protein Aggregates in Interferon-Alpha (IFN-α) Formulations [Elektronische Daten] [Andrea Braun, Jochem Alsenz] Purpose. Protein aggregates are thought to be involved in the immunogenicity of recombinant proteins in humans. To probe human IFN-α formulations for the presence of soluble protein aggregates, enzyme-linked immunosorbent assays (ELISA) were developed. Methods. For the detection of IFN-α-IFN-α and HSA-IFN-α aggregates, sandwich ELISAs were developed using a monoclonal anti-IFN-α antibody as a capture antibody and the same anti-IFN-α antibody and an anti-human serum albumin (HSA) antibody (HRP-labeled), respectively. Results. Marketed freeze-dried, HSA-containing IFN-α formulations tested in the ELISAs all contained IFN-α-IFN-α and/or HSA-IFN-α protein aggregates, although in varying amounts. These aggregates were predominantly IFN-α dimers and 1:1 conjugates of HSA with IFN-α. Test formulations revealed that aggregation of IFN-α was strongly affected by the presence of pharmaceutical excipients, pH of the formulation, lyophilisation procedure, and storage temperature and time. Conclusions. The ELISAs are rapid, highly specific for aggregates in the presence of both IFN-α and HSA monomers and allow the direct detection of both types of aggregates in formulations in the nanogram range. The new assays will assist the monitoring of the aggregate-inducing processes during IFN-α formulation and storage in an early phase and the development of aggregate-free IFN-α formulations. Plenum Publishing Corporation, 1997 interferon alpha (IFN-α) nationallicence human serum albumin (HSA) nationallicence enzyme-linked immunosorbent assay (ELISA) nationallicence protein formulation nationallicence protein aggregates nationallicence Braun Andrea Preclinical Research Department, Pharma Division, F. Hoffmann-La Roche Ltd, Grenzacherstrasse, CH-4002, Basel, Switzerland aut Alsenz Jochem Preclinical Research Department, Pharma Division, F. Hoffmann-La Roche Ltd, Grenzacherstrasse, CH-4002, Basel, Switzerland aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/10(1997-10-01), 1394-1400 0724-8741 14:10<1394 1997 14 11095 https://doi.org/10.1023/A:1012168621337 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012168621337 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Braun Andrea Preclinical Research Department, Pharma Division, F. Hoffmann-La Roche Ltd, Grenzacherstrasse, CH-4002, Basel, Switzerland aut NATIONALLICENCE 700 1- Alsenz Jochem Preclinical Research Department, Pharma Division, F. Hoffmann-La Roche Ltd, Grenzacherstrasse, CH-4002, Basel, Switzerland aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/10(1997-10-01), 1394-1400 0724-8741 14:10<1394 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer