Details
| Original language | English |
|---|---|
| Pages (from-to) | 177-183 |
| Number of pages | 7 |
| Journal | Analytica chimica acta |
| Volume | 374 |
| Issue number | 2-3 |
| Publication status | Published - 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.
Keywords
- Bioprocess monitoring, Flow-injection immunoassay (FIIA), Mammalian cell cultivation, OPA derivatization, rt-PA
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Environmental Science(all)
- Environmental Chemistry
- Chemistry(all)
- Spectroscopy
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Analytica chimica acta, Vol. 374, No. 2-3, 23.11.1998, p. 177-183.
Research output: Contribution to journal › Article › Research › 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 -