Match!

Application of Empirical Phase Diagrams for Multidimensional Data Visualization of High-Throughput Microbatch Crystallization Experiments

Published on Aug 1, 2018in Journal of Pharmaceutical Sciences3.197
· DOI :10.1016/j.xphs.2018.04.018
Marieke E. Klijn1
Estimated H-index: 1
(KIT: Karlsruhe Institute of Technology),
Jürgen Hubbuch32
Estimated H-index: 32
(KIT: Karlsruhe Institute of Technology)
Sources
Abstract
Abstract Protein phase diagrams are a tool to investigate the cause and consequence of solution conditions on protein phase behavior. The effects are scored according to aggregation morphologies such as crystals or amorphous precipitates. Solution conditions affect morphologic features, such as crystal size, as well as kinetic features, such as crystal growth time. Commonly used data visualization techniques include individual line graphs or phase diagrams based on symbols. These techniques have limitations in terms of handling large data sets, comprehensiveness or completeness. To eliminate these limitations, morphologic and kinetic features obtained from crystallization images generated with high throughput microbatch experiments have been visualized with radar charts in combination with the empirical phase diagram method. Morphologic features (crystal size, shape, and number, as well as precipitate size) and kinetic features (crystal and precipitate onset and growth time) are extracted for 768 solutions with varying chicken egg white lysozyme concentration, salt type, ionic strength, and pH. Image-based aggregation morphology and kinetic features were compiled into a single and easily interpretable figure, thereby showing that the empirical phase diagram method can support high-throughput crystallization experiments in its data amount as well as its data complexity.
  • References (69)
  • Citations (3)
📖 Papers frequently viewed together
1 Citations
1 Citations
21 Citations
78% of Scinapse members use related papers. After signing in, all features are FREE.
References69
Newest
#1Marie Therese Schermeyer (KIT: Karlsruhe Institute of Technology)H-Index: 1
#2Anna Katharina Wöll (KIT: Karlsruhe Institute of Technology)H-Index: 2
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 5 authors...
ABSTRACTHigh protein titers are gaining importance in biopharmaceutical industry. A major challenge in the development of highly concentrated mAb solutions is their long-term stability and often incalculable viscosity. The complexity of the molecule itself, as well as the various molecular interactions, make it difficult to describe their solution behavior. To study the formulation stability, long- and short-range interactions and the formation of complex network structures have to be taken into...
13 CitationsSource
#1Lara Galm (KIT: Karlsruhe Institute of Technology)H-Index: 6
#2Sven Amrhein (KIT: Karlsruhe Institute of Technology)H-Index: 5
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 3 authors...
The aggregation of proteins became one of the major challenges in the development of biopharmaceu­ticals since the formation of aggregates can affect drug quality and immunogenicity. However, aggregation mechanisms are highly complex and the investigation requires cost, time, and material intensive experi­mental effort. In the present work, the predictive power of protein characteristics for the phase behavior of three different proteins which are very similar in size and structure was studied. ...
10 CitationsSource
#1Halil I. Okur (EPFL: École Polytechnique Fédérale de Lausanne)H-Index: 12
#2Jana Hladílková (CAS: Academy of Sciences of the Czech Republic)H-Index: 5
Last. Pavel JungwirthH-Index: 55
view all 8 authors...
Ions differ in their ability to salt out proteins from solution as expressed in the lyotropic or Hofmeister series of cations and anions. Since its first formulation in 1888, this series has been invoked in a plethora of effects, going beyond the original salting out/salting in idea to include enzyme activities and the crystallization of proteins, as well as to processes not involving proteins like ion exchange, the surface tension of electrolytes, or bubble coalescence. Although it has been cle...
79 CitationsSource
#1Josefine Morgenstern (KIT: Karlsruhe Institute of Technology)H-Index: 5
#2Pascal Baumann (KIT: Karlsruhe Institute of Technology)H-Index: 8
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 4 authors...
Abstract During production, purification, formulation, and storage proteins for pharmaceutical or biotechnological applications face solution conditions that are unfavorable for their stability. Such harmful conditions include extreme pH changes, high ionic strengths or elevated temperatures. The characterization of the main influencing factors promoting undesired changes of protein conformation and aggregation, as well as the manipulation and selective control of protein stabilities are crucial...
17 CitationsSource
#1Katharina Christin Bauer (KIT: Karlsruhe Institute of Technology)H-Index: 7
#2Mathias Göbel (KIT: Karlsruhe Institute of Technology)H-Index: 1
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 5 authors...
The colloidal stability of a protein solution during downstream processing, formulation, and storage is a key issue for the biopharmaceutical production process. Thus, knowledge about colloidal solution characteristics, such as the tendency to form aggregates or high viscosity, at various processing conditions is of interest. This work correlates changes in the apparent diffusion coefficient as a parameter of protein interactions with observed protein aggregation and dynamic viscosity of the res...
12 CitationsSource
#1Kai Baumgartner (KIT: Karlsruhe Institute of Technology)H-Index: 5
#2Steffen Großhans (KIT: Karlsruhe Institute of Technology)H-Index: 4
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 5 authors...
Abstract In the biopharmaceutical industry it is mandatory to know and ensure the correct protein phase state as a critical quality attribute in every process step. Unwanted protein precipitation or crystallization can lead to column, pipe or filter blocking. In formulation, the formation of aggregates can even be lethal when injected into the patient. The typical methodology to illustrate protein phase states is the generation of protein phase diagrams. Commonly, protein phase behavior is shown...
6 CitationsSource
#1Marie-Therese Schermeyer (KIT: Karlsruhe Institute of Technology)H-Index: 3
#2Heike Sigloch (KIT: Karlsruhe Institute of Technology)H-Index: 2
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 5 authors...
This study aims at defining rheological parameters for the characterization of highly concentrated protein solutions. As a basis for comparing rheological behavior with protein solution characteristics the protein phase behavior of Lysozyme from chicken egg white with concentrations up to 225 mg/mL, changing pH values and additive concentrations was studied in a microbatch scale format. The prepared phase diagrams, scored after 40 days (t40) give insights into the kind and kinetics of the phase ...
10 CitationsSource
#1Lara Galm (KIT: Karlsruhe Institute of Technology)H-Index: 6
#2Josefine Morgenstern (KIT: Karlsruhe Institute of Technology)H-Index: 5
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 3 authors...
Abstract Undesired protein aggregation in general and non-native protein aggregation in particular need to be inhibited during bio-pharmaceutical processing to ensure patient safety and to maintain product activity. In this work the potency of different additives, namely glycerol, PEG 1000, and glycine, to prevent lysozyme aggregation and selectively manipulate lysozyme phase behavior was investigated. The results revealed a strong pH dependency of the additive impact on lysozyme phase behavior,...
12 CitationsSource
Non-native protein aggregation is common in the biopharmaceutical industry and potentially jeopardizes product shelf life, therapeutic efficacy, and patient safety. The present article focuses on the relationship(s) among protein–protein interactions, aggregate growth mechanisms, aggregate morphologies, and specific-ion effects for an anti-streptavidin (AS) immunoglobulin gamma 1 (IgG1). Aggregation mechanisms of AS-IgG1 were determined as a function of pH and NaCl concentration with sodium acet...
31 CitationsSource
#1Hauke Huettmann (Boehringer Ingelheim)H-Index: 2
#2Susanne Zich (BOKU: University of Natural Resources and Life Sciences, Vienna)H-Index: 1
Last. Alois Jungbauer (BOKU: University of Natural Resources and Life Sciences, Vienna)H-Index: 50
view all 5 authors...
Abstract Phase diagrams and solubility curves for engineering of crystallization for interferon gamma have been developed by small scale precipitation and crystallization experiments. A scale up was carried out to proof the accuracy and the suitability of the created phase diagram with sodium sulfate for bulk crystallization. To provide nucleation and a high yield the operating point was chosen at a protein concentration of 6 mg/mL and a sodium sulfate concentration of 25% [w/v]. Crystallization...
7 CitationsSource
Cited By3
Newest
#1Anna Katharina Wöll (KIT: Karlsruhe Institute of Technology)H-Index: 2
#2Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
Formulation conditions have a significant influence on the degree of freeze/thaw (FT) stress-induced protein instabilities. Adding cryoprotectants might stabilize the induced FT stress instabilities. However, a simple preservation of protein stability might be insufficient and further methods are necessary. This study aims to evaluate the addition of a heat cycle following FT application as a function of different cryoprotectants with lysozyme as exemplary protein. Sucrose and glycerol were show...
Source
#1Marieke E. Klijn (KIT: Karlsruhe Institute of Technology)H-Index: 1
#2Anna Katharina Wöll (KIT: Karlsruhe Institute of Technology)H-Index: 2
Last. Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
view all 3 authors...
Short-term parameters correlating to long-term protein stability, such as the protein cloud point temperature (Tcloud), are of interest to improve efficiency during protein product development. Such efficiency is reached if short-term parameters are obtained in a low volume and high-throughput (HT) manner. This study presents a low volume HT detection method for (sub-zero) Tcloud determination of lysozyme, as such an experimental method is not available yet. The setup consists of a cryogenic dev...
1 CitationsSource
#1Marieke E. Klijn (KIT: Karlsruhe Institute of Technology)H-Index: 1
#2Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
Abstract Image-based protein phase diagram analysis is key for understanding and exploiting protein phase behavior in the biopharmaceutical field. However, required data analysis has become a notorious time-consuming task since high-throughput screening approaches were implemented. A variety of computational tools have been developed to support analysis, but these tools primarily use end point visible light images. This study investigates the combined effect of end point and time-dependent image...
1 CitationsSource
#1Marieke E. Klijn (KIT: Karlsruhe Institute of Technology)H-Index: 1
#2Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
Abstract Redesigning existing food protein formulations is necessary in situations where food authorities propose dose adjustments or removal of currently employed additives. Redesigning formulations involves evaluating substitute additives to obtain similar long-term physical stability as the original formulation. Such formulation screening experiments benefit from comprehensive data visualization, understanding the effects of substitute additives on long-term physical stability, and identifica...
Source
#1Marieke E. Klijn (KIT: Karlsruhe Institute of Technology)H-Index: 1
#2Jürgen Hubbuch (KIT: Karlsruhe Institute of Technology)H-Index: 32
Abstract Identification of long-term stable biopharmaceutical formulations is essential for biopharmaceutical product development. Reduction of the number of long-term storage experiments and a well-defined formulation search space requires knowledge-based formulation screenings and a detailed protein phase behavior understanding. To achieve this, short-term analytical techniques can serve as predictors for long-term protein phase behavior. Protein phase behavior studies that investigate this co...
1 CitationsSource