themengebiete für studien- & abschlussarbeiten
TRANSCRIPT
KIT – University of the State of Baden-Wuerttemberg and
National Research Center of the Helmholtz Association
Institute of Process Engineering in Life Sciences
Section IV: Biomelecular Separation Engineering
www.kit.edu
Themengebiete für Studien- & Abschlussarbeiten
Doktoranden des MAB
Institute of Process Engineering in Life Sciences
Section IV: Biomolecular Separation Engineering
2
Allgemeines
Die folgende Übersicht dient dazu, Interessenten von Studien- bzw.
Abschlussarbeiten (BA, MA) einen Überblick über die Arbeitsgebiete
am Institut für „Molekulare Aufarbeitung von Bioprodukten“ zu geben.
Interessenten mit konkreten Themenwünschen können sich direkt bei
den jeweiligen Doktoranden melden oder allgemein bei David Grijalva
Themengebiete für Bachelor- und Masterarbeiten13.08.2021
3 13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
Angela Valentic
PhD project started 09/2019
Tasks: Analytical development + method development
Disassembly
and effect of
host cell nucleic
acids
Reassembly in
the presence of
nucleic acids
VLPVLP loaded
with RNA
Analytics:
- qPCR
- cGE
- SLS, DLS
- RP-/SEC- UHPLC
- Spectroscopic methods
Experimental:
- Process development + characterization of process
parameters by common and nucleic acid specific
analytical tools
- VLP production by cultivation in E.coli and purification by
e.g. precipitation, chromatography methods and UF/DF
@ThermoFisher@Wyatt @ThermoFisher
Changes in dispersity, structure and phase behaviour of
virus-like particles (VLPs) in the presence of nucleic acids
Motivation: Virus-like particles as carrier system for gene therapy
4
© ThermoFisher
Background
Carrier systems with nucleic acid cargo are
used for targeted delivery in the field of gene
therapy.
Robust production, purification and formula-
tion processes are required to ensure their
product quality attributes. Therefore, Process
Analytical Technology (PAT) is applied to
monitor process parameters by in-line, on-line
or off-line measurements.
13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
Annabelle Dietrich
Process development, analytical development and PAT implementation
for carrier systems with nucleic acid cargo.
© Spectrum Labs
Materials & Methods
Experiments- Process & analytical development
- Ultra-/Diafiltration (UF/DF)
- …
- PAT Implementation
Analytics - Spectroscopic methods (UV/Vis, Raman)
- Light Scattering
- RP-/SEC- UHPLC
- …
Tools- Matlab © Malvern
Virus-like particle (VLP)
Lipid-nanoparticle (LNP)
© TECAN
5
Christina Wegner – Protein Crystallization
Purification of biopharmaceuticals by protein crystallization is an attractive alternative to
chromatography. Up to now, laborious, empirical screenings are the core approach for
process development of crystallization processes. The combination of high throughput
screenings (HTS), spectroscopy, rheometry and elaborate data analysis can overcome
these limitations.
13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
PredictionPhase behaviour
On-line
analytics
HT screening
Purity
Yield
𝑐𝑝𝑟𝑜𝑡𝑒𝑖𝑛
Spectroscopy
𝑐𝑝𝑟𝑜𝑡𝑒𝑖𝑛
𝑐𝑐𝑜𝑛𝑡𝑎𝑚𝑖𝑛𝑎𝑛𝑡
Rheometry
𝜔𝐶𝑂
𝐺′
𝐺′′
Current projects:
- Process Analytical Technology (PAT) of solid and liquid
phase in lab-scale selective crystallization processes
- Prediction of crystallization behavior by protein-protein
interaction analysis
- Selective precipitation process development with
HTS of proteins in harvest broth
Methods:
- Spectroscopic methods (UV/Vis, Raman)
- Variable path length UV/Vis spectroscopy
- High frequency rheometry
- Multivariate data analysis in Matlab
- Analytical chromatography
- Pipetting roboter, automated screenings …
© Formulatrix
ddw-online.com
Dionex
Matlab
6 13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
David Grijalva – 3D Bioprinting
Background:Automated, reproducible manufacturing and analysis of
complex structures with biomaterials containing viable
cells
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eca
n
© A
nto
n-P
aa
rAnalytics:- Rheometry
- Flow cytometry
- Microscopy
- UV/Vis spectroscopy
Experimental:- Evaluation of biomaterials for 3D printing
applications
- Characterisation cell metabolic activities
© M
ilte
nyiB
iote
c
© Z
eis
s
7
Dennis Weber
Themengebiete für Bachelor- und Masterarbeiten13.08.2021
PROTEIN STABILITY IN FREEZE-THAW OPERATIONS
Material & Methods
Experiments
• µL- and mL cryodevices
• Robotic liquid handling station
Analytics
• SLS, DLS
• Melting temperature
• FT-IR
• …
CFD simulations
Background
Freezing and thawing are necessary
process units during the production of
biopharmaceuticals.
During the freezing process, freeze
concentration might occur. This may
lead to:
• Protein aggregation
• Surface denaturation
• pH denaturation
• Cold denaturation
• …
It is the aim of my thesis to evaluate the
protein stability during freezing/thawing.
ice
T / °
Cr / m
© T
ecan
© G
rant
© M
alv
ern
8 Themengebiete für Bachelor- und Masterarbeiten13.08.2021
Synthesis of Hybrid Nanoparticles via Aerosol Thiol-ene Photopolymerization
Gold Nanoparticle
HS
HS SH
3-functional ene
Aerosol
DropletsSpray SolutionPolymer
Nanoparticles
Atomization
Bio-agent
BioconjugationPhotoreaction
Bioconjugated
Nanoparticles
Narmin Suvarli
Legend:HS
HS SH
3-functional thiol Photoinitiator
Photothermal
therapy
Imaging
and Diagnostics
Pro
cess
Ap
pli
cati
on
s
• Cancer Screening and
treatment
• Drug delivery
• Encapsulation into hydrogels
for enzymatic reactions
Cancer tumor
Horizon 2020
Marie Sklodowska-Curie Actions
Innovative training network
9 Themengebiete für Bachelor- und Masterarbeiten13.08.2021
Nils Hillebrandt
Entwicklung eines hochdurchsatzfähigen Hochfrequenz-Rheometers zur Analyse der viskoelastischen Eigenschaften hochkonzentrierter Proteinlösungen [1]
[1] Kooperation mit dem Institut für Automation und angewandte Informatik (IAI), Ansprechpartner: Prof. Dr. Christian Pylatiuk
[2] Schermeyer et al. (2016). Squeeze flow rheometry as a novel tool for the characterization of highly concentrated protein solutions.
Biotechnology and Bioengineering, 113(3), 576–587. https://doi.org/10.1002/bit.25834
Ziel
Korrelation viskoelastischer
Eigenschaften mit der Lösungsstabilität
von Proteinlösungen [2]
Aufgaben
Evaluierung und konzeptionelle
Optimierung von
Messaufbau/Konstruktion
Design und Implementierung der
Automatisierungs- und
Temperierungseinheit
Techniken/Arbeitsbereiche
Ansteuerung von
piezokeramischen Aktoren und
Sensoren
Schwingungsanalyse
Programmierung mit MATLAB
Ansteuerung der peripheren
Bauteile (Pumpen, Peltier-
Element, Temperatur-
Controller
10 Themengebiete für Bachelor- und Masterarbeiten13.08.2021
Advancingultra- and
diafiltration
Online-Monitoring
ModelingHigh-
Throughput
Source:
https://www.sartorius.com/en/products/
process-filtration-purification/process-
filtration-hardware/crossflow-holders-
systems/crossflow-systems/ambr-
crossflow
Source: https://gosilico.com/
In collaboration with
Source:
http://spectrumlabs.com/filtration/K
R2System.html
Nils Hillebrandt
11 Themengebiete für Bachelor- und Masterarbeiten13.08.2021
Sandra Haas
Development and establishment of bioinks made of specially developed
elastic proteins for 3D printing of high-performance materials
Theoretical backgroundElastic materials are mostly petroleum-based and not biodegradable. The goal of my work is the use of biotechnologically
produced, elastin-like proteins (ELP) to produce highly elastic, 3D-printed hydrogels. Therefore, it is necessary to develop an
understanding of the factors influencing the later mechanical properties of the resulting hydrogels.
Ink developmentElastin-like proteins (ELP) 3D-Printing
• Development of protein analytics
• Optimisation of downstream processes
• Formulation of bioinks (model proteins and ELPs)
• Characterisation of formulated bioinks and hydrogels
• 3D printing using different techniques
(Stereolithography, Extrusion)
12 13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
Background:Automated, reproducible
manufacturing and analysis of
complex structures with biomaterials
containing viable cells
© T
eca
n
© A
nto
n-P
aa
r
Analytics and tools:- Rheometry
- Microscopy
- Image processing
- Matlab
Experimental:- Printing optimisation of biomaterials
- Development of standardised analytical
methods
Svenja Strauß – 3D Bioprinting
13
Process modeling and control• PAT tools in combination with mechanistic
models for all steps in the conjugation workflow
• Model-predictive control of conjugation reaction
Experimental Methods:▪ Kinetics determination▪ Analytics: RP-HPLC▪ UV-Vis spectroscopy
CFD-based reactor modeling▪ Scale-down reactor modeling
▪ Hydrodynamics-guided process scale-up for ADC reaction
▪ Full-3D conjugation reaction model
Simulation Methods:▪ 0D-Simulations with MATLAB®▪ CFD Simulation with Ansys Fluent®
13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
Jan WeggenAdvanced methodologies for modelling antibody-drug conjugation
Background: ADC production comprise a set of stirred tank operations, coupled with UF/DFfor buffer exchange and at least one chromatographic step. To increase processunderstanding, enable trouble shooting and ease process characterization as well as scalebridging, a set-up such as a ‘Digital Twin’ is developed.
Antibody-Drug Conjugate (ADCs):
© W
ater
s co
rpo
rati
on
© D
ion
ex
Conjugation
HIC
UF/DF
#Digital Twin
𝑓(𝑥)
14 13.08.2021 Themengebiete für Bachelor- und Masterarbeiten
Robin Schiemer
Artificial Intelligence for Bioprocess Development
Artificial Intelligence (AI) and Machine Learning (ML) methods provide powerful toolboxes for advanced data analytics with the increasing computation power
enabling the use of these algorithms in all sorts of sciences. In bioprocesses, process variables are monitored by in-line or off-line measurements which carry
information about the progress of the process. Spectroscopic sensors such as UV/VIS, FTIR or Raman sensors were shown to be reliable sources of process
information concerning the concentration of the contained species, excipient concentrations or structural conformations of the components. Parallely,
mechanistic models were established for various unit operations to save experimental time and costs and considerably increase process understanding. AI/ML
methods can help to create hybrid modeling strategies where real-time process information is combined with model predictions to successfully implement
efficient and precise process control.
Process
Sensor
Data Science
Mechanistic Modeling
capturereal-time information
extract patterns& makeprediction
use process knowledge
makeprediction
model-basedcontrol
Materials & Methods
Process Analytical Technologies (PAT) and mechanistic models are
applied to improve gene therapy process development and enable
model-predictive control of monoclonal antibody purification processes.
Experimental:
» Chromatography et al.
» Spectroscopy (UV/VIS, FTIR, Raman)
» Light Scattering (Zetasizer, MALS, RI)
» Analytics (HPLC, qPCR, ..)
Computational:
» Data Science (Python, TensorFlow)
» Process automation (MATLAB)
» Mechanistic modeling (ChromX)