Towards Protein Crystallization as a Process Step in Downstream Processing of Therapeutic Antibodies: Screening and Optimization at Microbatch Scale

Crystallization conditions of an intact monoclonal IgG4 (immunoglobulin G, subclass 4) antibody were established in vapor diffusion mode by sparse matrix screening and subsequent optimization. The procedure was transferred to microbatch conditions and a phase diagram was built showing surprisingly low solubility of the antibody at equilibrium. With up-scaling to process scale in mind, purification efficiency of the crystallization step was investigated. Added model protein contaminants were excluded from the crystals to more than 95%. No measurable loss of Fc-binding activity was observed in the crystallized and redissolved antibody. Conditions could be adapted to crystallize the antibody directly from concentrated and diafiltrated cell culture supernatant, showing purification efficiency similar to that of Protein A chromatography. We conclude that crystallization has the potential to be included in downstream processing as a low-cost purification or formulation step

Rapid SDS-GEL capillary electrophoresis for the analysis of recombinant NADP + -dependent formate dehydrogenase during expression in Escherichia coli cells and its purification

Abstract The level of expression in Escherichia coli cells and different steps of purification of the recombinant NADP + -dependent formate dehydrogenase (EC 1.2.1.2, FDH) from bacterium Pseudomonas sp.101 was analyzed by rapid SDS-Gel capillary electrophoresis (SDS-Gel CE) and compared with SDS polyacrylamide gel electrophoresis (SDS PAGE). First standard proteins were separated in the short capillary and the calibration curve generated, then fractions taken during the fermentation and purification process were analyzed. The main advantages of SDS-Gel CE are short analysis time, high sensitivity, the possibility to quantify proteins at different ultraviolet wavelength, and small injection volumes. The data for each step of the fermentation process and during the purification were controlled by spectrophotometric analysis of enzyme activity and protein concentration as well as standard SDS PAGE. The molecular mass of the purified FDH was determined as 44 078 Da by matrix-assisted laser desorption/ ionisation time of flight mass spectrometry © 1997 Elsevier Science B.V

Improved high-performance liquid chromatographic method in the analysis of adenovirus particles

NRC publication: Ye

Continuous Bed HPLC in the detection and quantification of adenoviruses

NRC publication: Ye

Improved high-performance liquid chromatographic method in the analysis of adenovirus particles

NRC publication: Ye

Purification of a Fab'Fragment by expanded bed absorption chromatography

NRC publication: Ye

Metod farmakologicheskoy korrektsii metabolizma kostnoy tkani dlya uluchsheniya rezul'tatov endoprotezirovaniya tazobedrennogo sustava

Пособие для врачей посвящено улучшению результатов эндопротезирования пациентов, страдающих остеопорозом или другими заболеваниями, при которых нарушается ремоделирование костной ткани (болезнь Педжета, гиперпаратиреоидная остеодистрофия после удаления аденомы околощитовидной железы, остеомаляция). Разработанный способ фармакологической коррекции нарушений ремоделирования костной ткани защищен патентом (патент РФ на изобретение № 2176519 от 10.12.01) и основан на комбинированной лекарственной терапии с использованием кальцитонина (миакальцика применяемого интраназально или в виде внутримышечных инъекций), альфакальцидола (альфа-D3 ТЕВА, Этальфа оксидевита) и препаратов кальция. Приведены факторы риска асептической нестабильности эндопротезов и схемы профилактики и лечения в зависимости от степени риска ее развития. Метод предназначен для улучшения результатов эндопротезирования и реэндопротезирования тазобедренного сустава

Anything but Conventional Chromatography Approaches in Bioseparation

While packed bed chromatography, known as conventional chromatography, has been serving the biopharmaceutical industry for decades as the bioseparation method of choice, alternative approaches are likely to take an increasing leading role in the next few years. The high number of new biological drugs under development, and the need to make biopharmaceuticals widely accessible, has been driving the academia and industry in the quest of anything but conventional chromatography approaches. In this perspective paper, these alternative approaches are discussed in view of current and future challenges in the downstream processing field