PhD degree in Chemistry
Chemical Engineering or Materials Engineering fields (Completed within last four years). At least 2 years of PDF Experience Working knowledge of polymer synthesis (ex.
lipid-polymer conjugation through amide
urethane
ester and imine linkages). Must have experience with Polymerization techniques like RAFT polymerization
ring opening polymerization and click chemistry. Experience in working with different MW PEI (from 0.6 to 25k MW) and purification techniques is an asset. Purification of polymers (ex.
precipitation
dialysis
and freeze drying)
knowledge and experience in storage
stability and half life period of polymers. The ability to reproduce the polymers in a bulk scale is an asset. Hands on experience with characterization for the functionalized polymers (1H and 13C NMR
FTIR
and MALDI). From NMR
know how to calculate the DS
and correlate the DS with the theoretical substitution. In addition
measure pKa (TFS assay) and CMC. Knowledge of binary and ternary polyplex formulations (via electrostatic interaction) based on different w/w or N/P ratio between polymer and genetic materials (pDNA
mRNA
siRNA and CRISPR). Additionally
must have knowledge of using different types of additives to improve the ternary polyplex formulation. Characterization of polyplex formulation (binding capacity via gel electrophoresis or SYBR green assay and dissociation capacity through heparin displacement). Physiochemical characterizations like particle size
zeta potential
PDI
and TEM. Hands on experience with transfection (both normal and reverse transfection) of polyplex formulations in different cell lines (attachment and suspension cells) such as A549
skin fibroblast
MDA-231
436
HCT116
Bone marrow cells and Dorsal root ganglia (DRG) neurons
Jurkat T cells
K562 WT
K562 IMR
K562 GFP
MOLM 13
MV4;11 cells. Hands on experience with RBC and PBMCs separation from Human blood via Ficoll-Paque protocol. Knowledge of hemocompatibility assay is an asset. Hands on experience with cell cytotoxicity studies (cell count and MTT)
serum stability studies
cell uptake
uptake mechanism (using different inhibitors and lysotracker reagents) and cell transfection (should handle flow cytometry
plate reader
fluorescence microscope and confocal microscope). Working knowledge of corelate uptake and transfection with pKa
CMC
size
zeta potential
lipidic substitution
lipidic tail length
w/w ratio
MW of primary polymer. Also
know how to correlate the transfection between binary and ternary polyplex formulations. Working knowledge of pDNA
mRNA
siRNA and CRISPR technologies. Experience with in-vivo imaging and organs collection