Immunobased elution assay for process control

authored by: Kerstin Beyer, Martin Reinecke, Wolfgang Noe, Thomas Scheper
Abstract: A flow-injection analysis (FIA) system with an integrated immunobased elution assay was developed for the on-line determination of biotechnological products. This paper describes its application for tissue-type plasminogen activator (rt-PA) and recombinant antithrombin III (r-AT III). The short response time of the immunochemical system allows its use for measurement and control of biotechnological production processes. Immobilized polyclonal antibodies were found to bind rt-PA in the cartridge better than monoclonal antibodies. Sepharose-4B was found to be the best support for the antibodies. The cartridge with immobilized antibodies can be prepared easily, has a high reliability and is stable for more than two weeks. The total time necessary for one assay cycle is in the range of 6-8 min, depending on the conditions. The whole procedure is controlled by a personal computer. The FIA system can be used for automatic analysis as a stand-alone version with a frequency of 7 samples per hour. The standard operation range is from 500 μg/ml down to 2 μg/ml. A comparison with the conventional microtiter ELISA assay shows that this assay can be applied for an accurate determination of undiluted cultivation samples.
External Organisation(s): Boehringer Ingelheim
University of Münster
Type: Article
Journal: Analytica chimica acta
Volume: 309
Pages: 301-305
No. of pages: 5
ISSN: 0003-2670
Publication date: 20.06.1995
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Analytical Chemistry, Biochemistry, Environmental Chemistry, Spectroscopy
Electronic version(s): https://doi.org/10.1016/0003-2670(95)00092-E (Access: Unknown)

BibTeX

@article{703dc0eaf6ff49729fe0951b7b03dc94,
title = "Immunobased elution assay for process control",
abstract = "A flow-injection analysis (FIA) system with an integrated immunobased elution assay was developed for the on-line determination of biotechnological products. This paper describes its application for tissue-type plasminogen activator (rt-PA) and recombinant antithrombin III (r-AT III). The short response time of the immunochemical system allows its use for measurement and control of biotechnological production processes. Immobilized polyclonal antibodies were found to bind rt-PA in the cartridge better than monoclonal antibodies. Sepharose-4B was found to be the best support for the antibodies. The cartridge with immobilized antibodies can be prepared easily, has a high reliability and is stable for more than two weeks. The total time necessary for one assay cycle is in the range of 6-8 min, depending on the conditions. The whole procedure is controlled by a personal computer. The FIA system can be used for automatic analysis as a stand-alone version with a frequency of 7 samples per hour. The standard operation range is from 500 μg/ml down to 2 μg/ml. A comparison with the conventional microtiter ELISA assay shows that this assay can be applied for an accurate determination of undiluted cultivation samples.",
keywords = "Flow injection, Immunoassay, Process analysis",
author = "Kerstin Beyer and Martin Reinecke and Wolfgang Noe and Thomas Scheper",
year = "1995",
month = jun,
day = "20",
doi = "10.1016/0003-2670(95)00092-E",
language = "English",
volume = "309",
pages = "301--305",
journal = "Analytica chimica acta",
issn = "0003-2670",
publisher = "Elsevier",
number = "1-3",
}