Crystallization of recombinant human growth hormone at elevated pressures: Pressure effects on PEG‐induced volume exclusion interactions

Crisman, Ryan L.; Randolph, Theodore W.

doi:10.1002/bit.22832

Cited by 16 publications

(15 citation statements)

References 45 publications

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“…In previous experiments on the 6 mL‐scale, it was observed that the process was accelerated by addition of small amounts of PEG 10000 (data not shown). This confirmed observations reported in literature (Crisman and Randolph, ; Jion et al, ; Weber et al, ). Therefore, 2% w/v PEG 10000 was added to the pretreated harvests to accelerate the first crystallization steps.…”

Section: Resultssupporting

confidence: 93%

Fast and scalable purification of a therapeutic full‐length antibody based on process crystallization

Smejkal

Agrawal

Helk

et al. 2013

Biotech & Bioengineering

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The potential of process crystallization for purification of a therapeutic monoclonal IgG1 antibody was studied. The purified antibody was crystallized in non-agitated micro-batch experiments for the first time. A direct crystallization from clarified CHO cell culture harvest was inhibited by high salt concentrations. The salt concentration of the harvest was reduced by a simple pretreatment step. The crystallization process from pretreated harvest was successfully transferred to stirred tanks and scaled-up from the mL-scale to the 1 L-scale for the first time. The crystallization yield after 24 h was 88-90%. A high purity of 98.5% was reached after a single recrystallization step. A 17-fold host cell protein reduction was achieved and DNA content was reduced below the detection limit. High biological activity of the therapeutic antibody was maintained during the crystallization, dissolving, and recrystallization steps. Crystallization was also performed with impure solutions from intermediate steps of a standard monoclonal antibody purification process. It was shown that process crystallization has a strong potential to replace Protein A chromatography. Fast dissolution of the crystals was possible. Furthermore, it was shown that crystallization can be used as a concentrating step and can replace several ultra-/diafiltration steps. Molecular modeling suggested that a negative electrostatic region with interspersed exposed hydrophobic residues on the Fv domain of this antibody is responsible for the high crystallization propensity. As a result, process crystallization, following the identification of highly crystallizable antibodies using molecular modeling tools, can be recognized as an efficient, scalable, fast, and inexpensive alternative to key steps of a standard purification process for therapeutic antibodies.

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Section: Resultssupporting

confidence: 93%

Fast and scalable purification of a therapeutic full‐length antibody based on process crystallization

Smejkal

Agrawal

Helk

et al. 2013

Biotech & Bioengineering

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“…The minimum in J(p) around 150 MPa remains up to 400 mM NaCl, and the J-value increases with increasing NaCl concentration, i.e., with increased screening of the positively charged lysozyme (Fig. The observed pressure dependence of the second virial coefficient is in good agreement with observations by Crisman and Randolph (2010) who studied the crystallization behavior of a human growth hormone and found an increase of the virial coefficient with increasing pressure from 0.1 to 250 MPa. As shown in Fig.…”

Section: Salt Effects On Intermolecular Interactions Of Proteins At Hsupporting

confidence: 86%

“…The latter parameter was explored in a few studies only, such as on subtilisin, glucose isomerase, thaumatin, SNase, RNase A, and lysozyme (Webb et al 1999;Visuri et al 1990;Waghmare et al 2000;Crisman and Randolph 2010;Kadri et al 2003Kadri et al , 2005Jaenicke 1991, 1993;Schall et al 1994;Saikumar et al 1995;Lorber et al 1996;Takano et al 1997;Suzuki et al 2002aSuzuki et al ,b, 2005Nagatoshi et al 2003;Möller et al 2012Möller et al , 2014bJavid et al 2007a, b). The latter parameter was explored in a few studies only, such as on subtilisin, glucose isomerase, thaumatin, SNase, RNase A, and lysozyme (Webb et al 1999;Visuri et al 1990;Waghmare et al 2000;Crisman and Randolph 2010;Kadri et al 2003Kadri et al , 2005Jaenicke 1991, 1993;Schall et al 1994;Saikumar et al 1995;Lorber et al 1996;Takano et al 1997;Suzuki et al 2002aSuzuki et al ,b, 2005Nagatoshi et al 2003;Möller et al 2012Möller et al , 2014bJavid et al 2007a, b).…”

Section: Salt Effects On Intermolecular Interactions Of Proteins At Hmentioning

confidence: 99%

High Pressure Bioscience

Akasaka¹,

Matsuki²

2015

Subcellular Biochemistry

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The book series SUBCELLULAR BIOCHEMISTRY is a renowned and well recognized forum for disseminating advances of emerging topics in Cell Biology and related subjects. All volumes are edited by established scientists and the individual chapters are written by experts on the relevant topic. The individual chapters of each volume are fully citable and indexed in Medline/Pubmed to ensure maximum visibility of the work.

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“…Recombinant human growth hormone was expressed in Escherichia coli and purified as described previously. 8,26 HES (Viastarch) was purchased from Fresenius (Graz, Austria), and sucrose and trehalose were purchased from Mallinckrodt Baker (Phillipsburg, New Jersey). All other chemicals were purchased as reagent grade or higher.…”

Section: Methodsmentioning

confidence: 99%

Protein Quantity on the Air–Solid Interface Determines Degradation Rates of Human Growth Hormone in Lyophilized Samples

Grobelny

Allmen

et al. 2014

Journal of Pharmaceutical Sciences

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rhGH was lyophilized with various glass-forming stabilizers, employing cycles that incorporated various freezing and annealing procedures to manipulate glass formation kinetics, associated relaxation processes and glass specific surface areas (SSA’s). The secondary structure in the cake was monitored by IR and in reconstituted samples by CD. The rhGH concentrations on the surface of lyophilized powders were determined from ESCA. Tg, SSA’s and water contents were determined immediately after lyophilization. Lyophilized samples were incubated at 323 K for 16 weeks, and the resulting extents of rhGH aggregation, oxidation and deamidation were determined after rehydration. Water contents and Tg were independent of lyophilization process parameters. Compared to samples lyophilized after rapid freezing, rhGH in samples that had been annealed in frozen solids prior to drying, or annealed in glassy solids after secondary drying retained more native-like protein secondary structure, had a smaller fraction of the protein on the surface of the cake and exhibited lower levels of degradation during incubation. A simple kinetic model suggested that the differences in the extent of rhGH degradation during storage in the dried state between different formulations and processing methods could largely be ascribed to the associated levels of rhGH at the solid-air interface after lyophilization.

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Crystallization of recombinant human growth hormone at elevated pressures: Pressure effects on PEG‐induced volume exclusion interactions

Cited by 16 publications

References 45 publications

Fast and scalable purification of a therapeutic full‐length antibody based on process crystallization

Fast and scalable purification of a therapeutic full‐length antibody based on process crystallization

High Pressure Bioscience

Protein Quantity on the Air–Solid Interface Determines Degradation Rates of Human Growth Hormone in Lyophilized Samples

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