Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 177-183 |
| Seitenumfang | 7 |
| Fachzeitschrift | Analytica chimica acta |
| Jahrgang | 374 |
| Ausgabenummer | 2-3 |
| Publikationsstatus | Veröffentlicht - 23 Nov. 1998 |
Abstract
A flow-injection immunoassay (FIIA) using ortho-phthalaldehyde (OPA) derivatization was developed for on-line monitoring of protein products in biotechnological processes. Recombinant tissue-type plasminogen activator (rt-PA) and its antibodies were used in these studies as a model system. The derivatization of rt-PA with OPA led to a lower detection limit (higher fluorescence intensity) and an increase of sensitivity (higher slope of calibration curve) for rt-PA compared with the method without OPA derivatization. The derivatization of rt-PA with OPA was also systematically characterized: influence of pH, derivatization time, OPA concentration and sample volume on fluorescence intensity. Interferences caused by proteins were investigated with different washing buffer solutions. A 0.1M potassium phosphate buffer (PPB) solution containing 1gl-1 polyethylene glycol (PEG) was found to be optimal for minimizing the interference with BSA in the measurement. On-line monitoring for rt-PA is carried out on a simulated bioprocess in the range 0-20μgml-1. The data obtained showed good agreement with the conventional off-line enzyme linked immunosorbent assay (ELISA) data. The FIIA using o-phthalaldehyde (OPA) derivatization is a valuable tool for on-line monitoring of low concentrated protein products in biotechnical production processes. Copyright (C) 1998 Elsevier Science B.V.
ASJC Scopus Sachgebiete
- Chemie (insg.)
- Analytische Chemie
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biochemie
- Umweltwissenschaften (insg.)
- Umweltchemie
- Chemie (insg.)
- Spektroskopie
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in: Analytica chimica acta, Jahrgang 374, Nr. 2-3, 23.11.1998, S. 177-183.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Flow-injection immunoassay using OPA derivatization
AU - Rhee, Jong Il
AU - Hagedorn, Jörg
AU - Kretzmer, Gerlinde
AU - Scheper, Thomas
AU - Schügerl, Karl
PY - 1998/11/23
Y1 - 1998/11/23
N2 - A flow-injection immunoassay (FIIA) using ortho-phthalaldehyde (OPA) derivatization was developed for on-line monitoring of protein products in biotechnological processes. Recombinant tissue-type plasminogen activator (rt-PA) and its antibodies were used in these studies as a model system. The derivatization of rt-PA with OPA led to a lower detection limit (higher fluorescence intensity) and an increase of sensitivity (higher slope of calibration curve) for rt-PA compared with the method without OPA derivatization. The derivatization of rt-PA with OPA was also systematically characterized: influence of pH, derivatization time, OPA concentration and sample volume on fluorescence intensity. Interferences caused by proteins were investigated with different washing buffer solutions. A 0.1M potassium phosphate buffer (PPB) solution containing 1gl-1 polyethylene glycol (PEG) was found to be optimal for minimizing the interference with BSA in the measurement. On-line monitoring for rt-PA is carried out on a simulated bioprocess in the range 0-20μgml-1. The data obtained showed good agreement with the conventional off-line enzyme linked immunosorbent assay (ELISA) data. The FIIA using o-phthalaldehyde (OPA) derivatization is a valuable tool for on-line monitoring of low concentrated protein products in biotechnical production processes. Copyright (C) 1998 Elsevier Science B.V.
AB - A flow-injection immunoassay (FIIA) using ortho-phthalaldehyde (OPA) derivatization was developed for on-line monitoring of protein products in biotechnological processes. Recombinant tissue-type plasminogen activator (rt-PA) and its antibodies were used in these studies as a model system. The derivatization of rt-PA with OPA led to a lower detection limit (higher fluorescence intensity) and an increase of sensitivity (higher slope of calibration curve) for rt-PA compared with the method without OPA derivatization. The derivatization of rt-PA with OPA was also systematically characterized: influence of pH, derivatization time, OPA concentration and sample volume on fluorescence intensity. Interferences caused by proteins were investigated with different washing buffer solutions. A 0.1M potassium phosphate buffer (PPB) solution containing 1gl-1 polyethylene glycol (PEG) was found to be optimal for minimizing the interference with BSA in the measurement. On-line monitoring for rt-PA is carried out on a simulated bioprocess in the range 0-20μgml-1. The data obtained showed good agreement with the conventional off-line enzyme linked immunosorbent assay (ELISA) data. The FIIA using o-phthalaldehyde (OPA) derivatization is a valuable tool for on-line monitoring of low concentrated protein products in biotechnical production processes. Copyright (C) 1998 Elsevier Science B.V.
KW - Bioprocess monitoring
KW - Flow-injection immunoassay (FIIA)
KW - Mammalian cell cultivation
KW - OPA derivatization
KW - rt-PA
UR - http://www.scopus.com/inward/record.url?scp=0031766170&partnerID=8YFLogxK
U2 - 10.1016/S0003-2670(98)00467-X
DO - 10.1016/S0003-2670(98)00467-X
M3 - Article
AN - SCOPUS:0031766170
VL - 374
SP - 177
EP - 183
JO - Analytica chimica acta
JF - Analytica chimica acta
SN - 0003-2670
IS - 2-3
ER -