High‐throughput process development methods for chromatography and precipitation of proteins: Advantages and precautions. Issue 5 (23rd August 2013)

• Record Type: Journal Article
• Authors: Yoshimoto, Noriko; Minakuchi, Kazunobu; Itoh, Daisuke; Isakari, Yu; Yamamoto, Shuichi
• Is Part Of: Engineering in life sciences. Volume 13:Issue 5(2013:Sep.)
• Page Start: 446
• Page End: 455
• Publication Date: 2013-08-23
• Subjects: Bioengineering -- Periodicals; 660.605
• DOI: 10.1002/elsc.201200121
• Languages: English
• ISSNs: 1618-0240

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Although high‐throughput process development methods using 96‐well microplate (MP) systems are promising and attractive, they still have several unclear problems in the practical applications. Several case studies for the purification process development by using manually operated 96‐well MP systems are presented. In the first example, the MPs with ion‐exchange monolith disks or ion‐exchange membranes were employed for linear gradient elution and breakthrough curve experiments. The data such as the peak salt concentration and dynamic binding capacity values obtained with the MP system were in good agreement with those by the LC system. In the second example, batch adsorption experiments with a 96‐well filter MP were performed for a protein A resin (particle)–antibody system in order to obtain the adsorption isotherms. The static binding capacities calculated from the isotherms were similar to those calculated from the LC system. In the last example, protein precipitation experiments by PEG were carried out for determining the solubility curve. It was found that the presence of dimer and aggregates affect the solubility curve measurement significantly. The proposed methods suggest powerful practical applications of high‐throughput process development for chromatography and precipitation process when the experiments are carefully designed and carried out.</p> </abstract>