Chapter Four · failure evidence
What Lipid & Membrane Engineering got wrong, from 71 dissertations
The records document pervasive engineering bottlenecks across the development, stabilization, and application of lipid formulations and artificial membranes. Major obstacles include membrane protein denaturation during detergent extraction, nanoparticle aggregation, low cytoplasmic delivery relative to simpler benchmarks, and physical artifacts in planar supported bilayers. These records come from PhD theses at 21 institutions, 2021 to 2026. Each links to its thesis. They were extracted by language models reading the full text, so treat each as a lead to read, not a verdict.
Detergent and polymer extraction destabilizes membrane proteins and disrupts native complexes
Extraction with detergents or synthetic polymers often causes membrane protein aggregation, unfolding, and non-specific co-purification of contaminants. These solubilization reagents frequently disrupt native protein-lipid interactions and produce large background micelle signals that hinder structural characterization.
Considered and rejected
Considered and rejected: Rejected detergent biochemical purification and nanodisc reconstitution of GPCRs due to delipidation, conformation loss, endogenous protein contamination, and lack of orientation control.
VIRION DISPLAY UNCOVERS MD-1 AS THE ENDOGENOUS AGONIST FOR THE ORPHAN RECEPTOR GPRC5B · JScholarship
Tried and failed
Detergent solubilisation for membrane protein cryo-EM applied to large bacterial membrane protein complexes. Reason: Large detergent micelle shells occluded structural features and produced substantial cryo-EM background noise
Investigating the protein subcomplexes from a conjugative Type IV Secretion System · Imperial
Tried and failed
nanodisc polymer membrane extraction applied to bacterial outer membrane lipoproteins. Reason: Signal peptidase cleavage yielded soluble truncated proteoforms instead of nanodisc-bound lipidated protein
Tried and failed
detergent or polymer solubilisation for particle purification applied to membrane-embedded protein pore complexes. Reason: failed to yield homogeneous pore particles during extraction from liposomes
Tried and failed
detergent solubilization for membrane protein purification applied to recombinant ligand-gated ion channels. Outcome: unstable. Reason: detergents caused protein aggregation and unfolding during size-exclusion chromatography
Biochemical, structural, and functional studies on the E. chrysanthemi ligand-gated ion channel · Texas Tech
Tried and failed
non-ionic detergent solubilization and affinity purification applied to integral membrane glycosyltransferases. Reason: detergent co-purified abundant non-specific endogenous host membrane protein contaminants
Biochemical Characterization of Glycan Assembly Pathway Enzymes · MIT
Tried and failed
non-ionic detergent lysis for single-molecule pulldown applied to membrane-associated protein complexes. Outcome: no signal. Reason: detergent solubilization disrupted native membrane protein interactions, preventing complex detection
Untangling Par complex dynamics during embryogenesis using single-molecule TIRF microscopy · UT Austin
Tried and failed
rapid dilution with bio-beads for proteoliposome reconstitution applied to purified membrane proteins. Outcome: unstable. Reason: rapid detergent removal caused protein aggregation and precipitation during liposome incorporation
Biochemical, structural, and functional studies on the E. chrysanthemi ligand-gated ion channel · Texas Tech
Tried and failed
mild non-ionic detergent solubilisation for co-fractionation mass spectrometry applied to membrane-associated protein complexes. Outcome: no signal. Reason: mild non-ionic detergents failed to extract and systematically depleted structural cytoskeletal actin proteins
The protein organization of a red blood cell · UT Austin
Tried and failed
styrene-maleic acid copolymer solubilization into nanodiscs applied to large monotopic membrane proteins. Reason: polymer failed to extract and solubilize the membrane proteins from lipid membranes
Biochemical Characterization of Glycan Assembly Pathway Enzymes · MIT
Tried and failed
membrane protein solubilization with harsh non-ionic detergents applied to transmembrane palmitoyltransferase enzyme complexes. Outcome: unstable. Reason: detergents Triton X-100, C9G, and LDAO induced complete protein aggregation
STRUCTURAL AND FUNCTIONAL STUDIES OF THE DHHC9 SUBFAMILY OF PROTEIN ACYLTRANSFERASES · Cornell
Tried and failed
nanodisc incorporation for solution NMR applied to membrane pore protein complexes. Reason: insufficient yield and poor sample homogeneity prevented 3D solution NMR structural studies
Biophysical studies of biological nanopores used for biopolymer sequencing · Imperial
Tried and failed
multiple pendant detergents and amphipols for solubilisation applied to membrane transport protein extraction. Outcome: unstable. Reason: failed to efficiently extract and stabilize the protein, causing aggregation during size-exclusion chromatography
Structural and functional characterisation of the human vitamin c transporters · Imperial
Considered and rejected
Considered and rejected: Rejected SMA/DIBMA lipid nanodiscs in favor of traditional DDM detergent because polymer nanodiscs previously had undesirable reagent interactions disrupting downstream assays.
Regulation of ABCG2 through interactions with Alpha-1-Acid Glycoprotein and EMMPRIN · University of Nottingham Repository
Considered and rejected
Considered and rejected: Standard alcoholic fixation and detergent permeabilization rejected for phosphoinositide microscopy because detergents disrupt membranes and extract PIs.
A lipidomic approach to elucidating the effects of antifungal compounds on signalling in fungi · Imperial
Considered and rejected
Considered and rejected: Rejected RIPA and Triton X-100 detergent lysis for organelle interactome pulldowns because detergents disrupt membrane integrity, labile protein interactions, and endosomal vesicles.
Activity-dependent activation of mTORC1 in neurons · JScholarship
Considered and rejected
Considered and rejected: Rejected detergent lysis buffer (deoxycholate) from PeriPreps kit in favor of water with Omnicleave endonuclease plus sonication to prevent solubilizing membrane proteins
The green fluorescent protein functions as a reporter for protein localization in Escherichia coli · Iowa State
Considered and rejected
Considered and rejected: Purifying solubilized GPCRs directly in detergent micelles away from membranes, due to protein unfolding, conformational instability, and MS incompatibility.
Elucidating the Role of Two Inotropic Agents in Cardiac Hypertrophy: the Ceramides and Apelins · ResearchWorks
Suboptimal lipid compositions and processing induce particle aggregation and colloidal instability
Lipid nanoparticles and liposomes lacking adequate steric stabilizers or formulated with long alkyl chains aggregate and exceed acceptable size thresholds. Mechanical processing, freeze-drying without cryoprotectants, and electrostatic screen failure in physiological ionic strengths similarly lead to membrane rupture and precipitation.
Tried and failed
incorporating cationic lipids in microfluidic lipid formulations applied to mRNA lipid nanoparticles. Outcome: unstable. Reason: formulations failed monodispersity and size quality control criteria and precipitated out of solution
Design and Formulation Optimization of Lipid Nanoparticles Carrying MRNA to the Heart and the Pancreas · Georgia Tech
Tried and failed
cationic lipid-mRNA complexes on mesoporous scaffolds applied to sustained in vivo mRNA delivery. Outcome: unstable. Reason: lipoplexes degraded rapidly at physiological temperature losing transfection capacity within days
Mesoporous Silica Rods Scaffolds for the generation of adaptive immune responses · Harvard
Tried and failed
formulation without steric stabilizer lipid applied to lipid nanoparticles. Outcome: unstable. Reason: aggregation and instability leading to particle diameter exceeding size quality control threshold (>200 nm)
UTILIZING HIGH-THROUGHPUT IN VIVO NANOPARTICLE SCREENS TO IMPROVE DRUG DELIVERY · Georgia Tech
Tried and failed
supported lipid bilayer coating applied to mesoporous silica microparticles. Outcome: unstable. Reason: low lipid-to-particle ratio caused particle aggregation
Materials to enable and enhance T-cell immunotherapies · Harvard
Tried and failed
nanoprecipitation with lipid-peg surfactant applied to hydrophobic conjugated polymers. Outcome: unstable. Reason: produced colloidally unstable nanoparticles with non-specific emission quenching
Modulating the properties of conjugated systems towards bioelectronics and bio-imaging · Imperial
Tried and failed
thin-film freeze-drying without lyoprotectant applied to lipid nanoparticle powder reconstitution. Outcome: unstable. Reason: absence of lyoprotectant caused vesicle damage and particle aggregation during freezing and drying
Lipid-based nanoparticles for drug delivery · UT Austin
Tried and failed
stearylamine-mediated surface charge modification of lipid nanoparticles applied to drug delivery nanocarriers. Outcome: unstable. Reason: formulations aggregated rapidly, yielded oversized particles, and exhibited high intrinsic cell cytotoxicity
In vitro functionality and toxicity of dacarbazine delivery nanosystem for melanoma. · Cranfield
Tried and failed
nanostructured lipid carrier emulsification solvent evaporation applied to chemotherapeutic drug delivery systems. Outcome: unstable. Reason: rapid particle re-aggregation within 72 hours, exceeding size limits and causing cytotoxicity
In vitro functionality and toxicity of dacarbazine delivery nanosystem for melanoma. · Cranfield
Tried and failed
cell membrane-coated nanoparticles without hydrophilic shielding polymer applied to intravenous nanoparticle delivery. Outcome: unstable. Reason: lack of protective surface coating caused particle aggregation and systemic toxicity in circulation
An apoptotic body-inspired nanoparticle to modulate inflammatory macrophages · UT Austin
Lost to a baseline
C16 alkyl chain ionizable lipids in the initial RNP LNP library resulted in particle aggregation (>1 µm size) and poor Cas9 protein encapsulation compared to C12 and C14 lipids.
Considered and rejected
Considered and rejected: Rejected NLC/GM and NLC/SI solid lipid formulations due to particle instability, aggregation, and unacceptable vehicle cytotoxicity.
In vitro functionality and toxicity of dacarbazine delivery nanosystem for melanoma. · Cranfield
Considered and rejected
Considered and rejected: Pivoted away from synthesizing ~1 um liposomes due to severe batch-to-batch size variability, opting for PLGA microparticles and targeted LNPs/hydrogels instead
Targeting the draining lymphatic network as a regulator of melanoma growth · Georgia Tech
Considered and rejected
Considered and rejected: Rejected DSPC, DOPE, and DOTAP formulations (100% lipid or with cholesterol/DSPE-PEG) for direct lipid coating of RENPs because they resulted in aggregation
Rare-Earth Nanoparticles for Non-invasive In Vivo Imaging of Immune Cells in Cancer Immunotherapy · MIT
Tried and failed
photoswitchable lipid bilayer formulation without membrane stabilizer applied to liposomal cargo retention at physiological temperature. Outcome: unstable. Reason: lack of cholesterol led to membrane destabilization and cargo leakage upon thermal incubation
LisNRs : a novel class of liposomal contrast agents for molecular MRI · MIT
Tried and failed
hydrophilic coating of polymer microfluidic channels applied to solvent-based liposome synthesis devices. Outcome: unstable. Reason: coatings suffered wash-out instability, causing channel wetting and lipid precipitation blockage
Developing artificial cells for delivering drug molecules to micro-engineered heart tissue · Imperial
Tried and failed
post-insertion of lipid-peg micelles applied to cholesterol-anchored aptamer liposomes. Outcome: unstable. Reason: micelle insertion displaced single-cholesterol anchored conjugates out of the bilayer into solution
Analysis of the properties of phospholipid bilayers for drug delivery using solid-state NMR · Imperial
Considered and rejected
Considered and rejected: Rejected evaluating liposomes below 100 nm diameter due to significantly decreased stability from elevated membrane surface energy/curvature.
Biomimetic water-based lubrication: an exploration into vesicle lubrication · Imperial
Tried and failed
monovalent charged lipid electrostatic stabilization applied to colloidal lipid nanodiscs. Outcome: unstable. Reason: electrostatic repulsion insufficient to maintain disc structure under physiological ionic strength conditions
Lipid nanoparticles frequently underperform simpler delivery baselines in transfection and cellular uptake
Formulations using permanent cationic lipids, neutral mixtures, or suboptimal charge ratios often fail to match the intracellular delivery of electroporation or peptide benchmarks. In vitro screening outcomes fail to correlate with in vivo efficacy, while high lipid ratios frequently introduce severe vehicle cytotoxicity.
Tried and failed
permanently charged cationic lipid substitution in nanoparticles applied to RNA delivery formulations. Outcome: worse than baseline. Reason: produced significantly lower transfection expression compared to degradable ionizable lipid benchmarks
Development of Chemically-Defined Platform Materials for Localized Delivery of RNA Therapeutics · MIT
Tried and failed
lipid nanoparticle delivery formulation applied to activated microglia mRNA transfection. Outcome: worse than baseline. Reason: inflammatory activation caused decreased delivery efficacy relative to basal state
Development of Chemically-Defined Platform Materials for Localized Delivery of RNA Therapeutics · MIT
Tried and failed
lowering cationic lipid to RNA ratio applied to lipid nanoparticle nucleic acid delivery. Outcome: worse than baseline. Reason: lowest N/P ratio nanoparticles unexpectedly showed higher cytotoxicity than intermediate ratios
Lipid-based nanoparticles for drug delivery · UT Austin
Tried and failed
intermediate charge ratio complexation in lipid nanoparticles applied to siRNA delivery and gene silencing. Outcome: worse than baseline. Reason: intermediate charge ratios exhibited slowest release kinetics, reducing downregulation efficacy
Lipid-based nanoparticles for drug delivery · UT Austin
Lost to a baseline
Cationic lipid library (e.g., X10-C12) and PEG-mediated RNP delivery achieved parity but failed to surpass control methods (TransIT-2020 and dried protein) in preliminary biolistic Cas9 RNP editing
Improving biolistic delivery for DNA and protein systems in plants · Iowa State University Digital Repository
Lost to a baseline
DSPC helper lipid formulation was outperformed up to four-fold in vivo by DOPE-containing LNPs for mRNA delivery
Engineering Lipid Nanoparticle-Mediated Delivery of Nucleic Acids to Lymphoid Organs and Immune Cells · DSpace at UTSWMED
Lost to a baseline
Peptide-only nanoparticles assembled manually or via Spark enabled higher DNA expression than all hybrid lipid-peptide formulations.
Use Of Microfluidic Systems For Assembly Of Nucleic Acid Nanoparticles · JScholarship
Lost to a baseline
In vivo delivery of Cas13-nLuc mRNA via MC3, Lipid 5, and LP01 LNPs resulted in lower liver luminescent flux compared to cKK-E12 LNPs.
Development of an mRNA-based prophylactic for protection against nerve agent toxicity · Georgia Tech
Tried and failed
in vitro screening to predict in vivo delivery applied to lipid nanoparticle nucleic acid formulations. Outcome: did not generalise. Reason: In vitro delivery efficacy showed no correlation with in vivo delivery across hundreds of formulation variants
UTILIZING HIGH-THROUGHPUT IN VIVO NANOPARTICLE SCREENS TO IMPROVE DRUG DELIVERY · Georgia Tech
Tried and failed
ternary uncharged lipid mixtures applied to intracellular chemotherapeutic drug delivery. Outcome: worse than baseline. Reason: insufficient interaction with cell surfaces due to lack of charged head groups
Engineering liposomal particles for direct, intracellular delivery of therapeutic molecules · UT Austin
Tried and failed
mTOR inhibitor pretreatment to enhance delivery applied to lipid nanoparticle siRNA delivery in vitro. Outcome: worse than baseline. Reason: mTOR inhibition failed to enhance silencing and impaired functional delivery at higher doses
The impact of the metabolic state of a cell on nucleic acid therapeutics · Georgia Tech
Lost to a baseline
At 100 µM doxorubicin, binary liposomes containing 3 mol% DOTAP killed only 8 ± 1% of cells compared to ~70% killed by free doxorubicin
Engineering liposomal particles for direct, intracellular delivery of therapeutic molecules · UT Austin
Lost to a baseline
Lipid nanoparticles (LNPs) achieved only 20.56% transfection efficiency in PCFs compared to 52.24% with electroporation.
Etablierung eines optogenetischen Systems zur lichtinduzierten Proteinbiosynthese in humane Stammzellen für therapeutische Anwendungen · Leibniz Universität Hannover Repository
Lost to a baseline
At 100 µM doxorubicin, unencapsulated (free) doxorubicin killed ~70% of cells, whereas ternary liposomes lacking DOTAP killed only 25% of cells.
Engineering liposomal particles for direct, intracellular delivery of therapeutic molecules · UT Austin
Considered and rejected
Considered and rejected: Rejected C3 LNP formulation at 20:1 N:P ratio despite higher in vitro delivery due to enhanced cytotoxicity and excess free ionizable lipid.
IONIZABLE LIPID NANOPARTICLES FOR IN UTERO GENE EDITING OF CONGENITAL DISEASE · Penn
Tried and failed
bilayer asymmetry engineering via inverted emulsion transfer applied to nanoparticle cellular uptake. Outcome: no signal. Reason: specific lipid compositions failed to mediate cellular internalization regardless of membrane asymmetry configuration
Considered and rejected
Considered and rejected: Rejected lipid/liposome-based transfection (e.g., FuGENE, Lipofectamine) for delivering CRISPR components into HUVECs due to high cytotoxicity, cytoplasmic nucleic acid degradation, and low efficiency compared to electroporation.
Dynamics of cytokine induced E-selectin expression in endothelial cells · University of Nottingham Repository
Planar and supported lipid bilayers introduce substrate artifacts and lack physiological curvature
Supported lipid bilayers and freestanding planar membranes exhibit severe edge defects, fluid instability, and substrate delamination under mechanical stress. These planar models also alter protein mobility and generate boundary artifacts in molecular dynamics that fail to mimic native vesicles.
Tried and failed
mechanical scratch patterning of thin surface coatings applied to supported lipid bilayer on conductive polymer. Outcome: unstable. Reason: scratching unintentionally delaminated the underlying soft polymer film along with the lipid bilayer
Lost to a baseline
Electrical sealing properties of NHC-based tBLMs (transmembrane resistance 177-194 kΩ/cm²) were inferior to the reference thiol anchor lipid DPhyTL (2.48-3 MΩ/cm²) due to membrane defect density
Moving in the right direction: Functionalized N-heterocyclic carbenes to assemble surface-bound biomolecular architectures for applications from biosensing to synthetic lipids · DSpace-CRIS at TU Wien
Considered and rejected
Considered and rejected: Rejected carboxy-methylated dextran (CM5) SPR chips for measuring physiological β3-talin affinity due to biphasic behavior, charge artefacts, and heterogeneous presentation compared to phospholipid bilayers
The Integrin Equilibrium: Balancing Protein-Protein and Protein-Lipid interactions · Penn
Considered and rejected
Considered and rejected: Rejected single-lased (1X-LIG) hydrophilic substrates for sensing because of water layer accumulation, membrane runoff over the stem, and large EMF drift.
Considered and rejected
Considered and rejected: Supported lipid bilayers and planar monolayers were avoided as primary models due to absence of bilayer curvature, substrate effects on lipid mobility, and vulnerability to air oxidation.
Second harmonic scattering of water in a biological context · EPFL
Considered and rejected
Considered and rejected: Rejected planar freestanding lipid bilayers as realistic cell membrane models because they are fragile, lack curvature, and naturally tend to close into vesicles to avoid exposed hydrophobic edges.
Tried and failed
small-angle neutron scattering without protein deuteration applied to detergent-solubilized membrane proteins. Outcome: no signal. Reason: hydrogenated protein signal could not be distinguished from detergent micelle scattering contributions
Tried and failed
homology crystal structure rigid-body SANS data fitting applied to membrane protein oligomers in bicelles. Outcome: did not converge. Reason: homolog crystal structure did not accurately represent the solution oligomeric conformation in lipid bicelles
Tried and failed
molecular dynamics of finite planar lipid bilayer applied to protein-membrane binding affinity simulation. Reason: Proteins preferentially adhered to hydrophobic periodic boundary or edge artifacts rather than the planar bilayer surface.
Characterizing & modelling the bio-nano interface · Imperial
Tried and failed
neutron spin echo on lipid bicelles applied to membrane protein dynamics. Outcome: no signal. Reason: the bicelle matrix was too fluidic, causing translational diffusion to dominate the internal dynamics signal
Assembly and processing conditions trigger payload leakage and nucleic acid degradation
Preparation methods such as sonication, extrusion, and flash nanoprecipitation cause substantial cargo loss, degradation of sensitive mRNA payloads, and reduced encapsulation efficiency. Suboptimal phase transition temperatures and cryoprotectant choices further exacerbate payload release or impair biological potency.
Tried and failed
nanoparticle formulation at low lipid to cargo ratio applied to lipid nanoparticles encapsulating nucleic acids. Outcome: worse than baseline. Reason: insufficient lipid mass ratio resulted in poor cargo encapsulation efficiency
Tried and failed
co-encapsulating disaccharide cryoprotectant during lipid nanoparticle assembly applied to polymer-coated lipid nanoparticle RNA delivery. Outcome: worse than baseline. Reason: sucrose co-encapsulation reduced transfection efficiency despite maintaining acceptable size and polydispersity
Considered and rejected
Considered and rejected: Rejected rehydration of dried lipid films in the presence of mRNA and extrusion due to complete loss/degradation of mRNA during sonication and extrusion.
Mesoporous Silica Rods Scaffolds for the generation of adaptive immune responses · Harvard
Tried and failed
lysolipid temperature-sensitive liposome formulation applied to thermally triggered vesicle payload release. Outcome: no signal. Reason: minimal cargo release occurred at target phase transition temperature despite formulation adjustments
Development of stimuli-responsive vesicles as distributed artificial organelles · Imperial
Tried and failed
lyophilization and reconstitution of functionalized liposomal formulations applied to therapeutic immunomodulatory protein-lipid nanoparticles. Outcome: worse than baseline. Reason: freeze-drying and reconstitution reduced target tumor cell lysis activity compared to freshly prepared formulations
Multivalent immunomodulators for CAR T cell manufacturing and T cell-engager immunotherapy · Georgia Tech
Tried and failed
direct liposome encapsulation of proteins applied to protein delivery vehicle preparation. Reason: low phase transition temperature caused cargo leakage, while higher transition temperature lipids caused thermal artifacts
Considered and rejected
Considered and rejected: Rejected microfluidic Tesla mixers and flash nanoprecipitation because Tesla mixers function primarily for lipid-polymeric hybrids and flash nanoprecipitation resulted in lower encapsulation efficiency.
Nanoparticle Fabrication via Microfluidics as a Therapeutic for Influenza A Virus · DSpace at SUNY Buffalo
Tried and failed
PEGylated lipid stabilization for protein encapsulation applied to lyotropic liquid crystalline nanoparticles. Outcome: worse than baseline. Reason: loaded significantly less protein cargo than poloxamer stabilization despite having larger internal water channels
Advanced single particle characterisation of cargo loading in therapeutic particles · Imperial
Considered and rejected
Considered and rejected: Rejected off-the-shelf cationic lipid formulations (e.g., DSTAP) or PEI for microbubble cargo loading due to unstable loading/enzymatic degradation and high polymer toxicity.
Microbubble-Assisted Ultrasound Guided Immunotherapy of Cancer: A Novel Microbubble Platform for Cancer Immunotherapy · DSpace at UTSWMED
Surface functionalization and helper lipid selection fail to redirect biodistribution away from liver clearance
Lipid formulations designed for cell targeting or pulmonary delivery often fail to achieve significant uptake in target tissues and remain sequestered in the liver or bone marrow. Incorporating antibody fragments, cationic cholesterol, or alternative ionizable lipids fails to improve delivery potency over simpler control vehicles.
Tried and failed
neutral or anionic lipid nanoparticle formulation applied to systemic mRNA delivery to lungs. Outcome: no signal. Reason: neutral and anionic helper lipids failed to facilitate measurable pulmonary biodistribution or transfection
Next-Generation Lipid Nanoparticle Formulations for Non-Liver Delivery of Nucleic Acid-Based Therapies and Vaccines · Georgia Tech
Lost to a baseline
Encapsulated rutin in lipid nanoparticles (LNPs) showed lower penetration (ART 16.3–16.5 MGV/px) than unencapsulated solutions in lemongrass oil (33.1 MGV/px) or MCT (29.1 MGV/px) in finite-dose assays
smartOX - Smarte Antioxidantien zur Pflege und Regeneration sensitiver Haut smartOX - Smart antioxidants for the care and regeneration of sensitive skin · open_UMR Marburg DSpace 10.0
Tried and failed
scFv targeting moieties instead of full antibodies applied to targeted lipid nanoparticle delivery. Outcome: worse than baseline. Reason: scFv conjugated nanoparticles failed to outperform full monoclonal antibody conjugates in delivery potency
Tried and failed
cationic lipid nanoparticle delivery applied to circulating blood cell targeting in vivo. Outcome: no signal. Reason: nanoparticles delivered cargo to liver and marrow but lacked statistically significant delivery to circulating blood cells
Considered and rejected
Considered and rejected: Rejected 1X antibody input mass during maleimide lipid conjugation because it failed to achieve targeted antibody densities per liposome
Multivalent immunomodulators for CAR T cell manufacturing and T cell-engager immunotherapy · Georgia Tech
Considered and rejected
Considered and rejected: LNPs formulated with proprietary ionizable lipids IL4-IL7 (LNP-017 through LNP-020) designed for extended circulation without antibody conjugation were rejected due to poor HSC targeting and preferential liver accumulation
Tried and failed
incorporating cationic cholesterol in lipid nanoparticles applied to intratumoral mRNA delivery. Outcome: worse than baseline. Reason: negatively enriched compared to neutral cholesterol formulations during in vivo screening
Translational Design of Lipid Nanoparticles (LNPs) to Deliver mRNA Therapies to Solid Tumors and to the Lungs · Georgia Tech
Membrane structural engineering compromises vesicle retention and enzymatic responsiveness
Omitting essential stabilizing lipids like cholesterol diminishes bilayer rigidity and causes cargo leakage, while artificial membrane cross-linking accelerates cargo loss. Sizing through extrusion or adding polyethylene glycol layers impairs outer shell hydration lubrication and reduces enzymatic responsiveness.
Lost to a baseline
Cross-linked liposome retention after 10 days (1.6% to 2.9%) was inferior to non-crosslinked standard liposomes (21.5% to 34.9%).
Pt(IV) anticancer prodrugs and liposomal encapsulation · Oxford
Considered and rejected
Considered and rejected: Rejected Ni-NTA and Co-NTA liposome formulations due to lack of stable binding of his-tagged peptides and antigens in biological media.
Developing a Novel Liposomal Adjuvant for Malaria Transmission-Blocking Vaccines · DSpace at SUNY Buffalo
Considered and rejected
Considered and rejected: Rejected preparing liposomes containing only phospholipids without cholesterol because cholesterol was required to increase bilayer rigidity, enhance solute retention, and expose maleimide residues for antibody coupling
Endothelial-specific targeting of Rho kinase signaling using CD31 antibody-conjugated nanoparticles · Texas Tech
Considered and rejected
Considered and rejected: Rejected PEGylation of liposomes because attached polyethylene glycol chains impede outer shell hydration lubrication.
Biomimetic water-based lubrication: an exploration into vesicle lubrication · Imperial
Considered and rejected
Considered and rejected: Rejected using 0.2 µm extruded liposomes in favor of polydisperse liposomes because extrusion reduced enzymatic cleavage sensitivity.
Activatable Fluorophores for the Molecular Imaging of Cytosolic Phospholipase A2 in Breast Cancer · Penn
Left open by the authors
Problems the authors named and did not get to.
Left open
Synthesize and formulate ionisable polymer-functionalised lipid nanoparticles for RNA delivery. Blocker: Requires a wet-lab environment and chemical/biological materials for nanoparticle synthesis and extrusion
Left open
Develop lipid nanoparticle-based mRNA delivery strategies for Cas9-retron genome editing systems. Blocker: Requires a wet lab for lipid nanoparticle formulation, mRNA synthesis, and in vitro/in vivo delivery assays
Discovery of novel systems for genome editing · UT Austin
Left open
Incorporate mRNAs containing modified UTRs into lipid nanoparticles to decrease off-target liver expression. Blocker: Requires wet-lab synthesis of modified mRNAs, formulation of lipid nanoparticles, and in vivo/in vitro delivery assays.
Left open
Modify the surface of polymer-functionalised lipid nanoparticles with specific targeting ligands for targeted RNA delivery. Blocker: Requires wet lab facilities for chemical synthesis, lipid nanoparticle formulation, and targeting assays
Left open
Evaluate how RNA design and LNP formulations impact antigen and inflammatory kinetics in mRNA vaccines. Blocker: Requires a wet lab to formulate lipid nanoparticles (LNPs), design RNA sequences, and measure in vivo or in vitro kinetics.
Left open
Test therapeutically relevant nucleic acid payloads formulated in lipid nanoparticles in an in vivo atherosclerosis mouse model. Blocker: Requires wet lab facilities, microfluidic LNP formulation, and in vivo animal disease models
Design and Formulation Optimization of Lipid Nanoparticles Carrying MRNA to the Heart and the Pancreas · Georgia Tech
Left open
Deliver therapeutic mRNA including CRISPR/Cas9 to the brain using lipid nanoparticles. Blocker: Requires a wet lab, chemical synthesis, microfluidic equipment, and in vivo animal models for brain delivery.
Development of Chemically-Defined Platform Materials for Localized Delivery of RNA Therapeutics · MIT
Left open
Deliver base and prime editors encapsulated in lipid nanoparticles to the lung endothelium in vivo. Blocker: Requires wet lab facilities for LNP formulation and animal models for in vivo delivery assays
Left open
Evaluate mRNA delivery by barcoded lipid nanoparticles to intratumoral immune cell subtypes in immunocompetent mice with spontaneous tumors. Blocker: Requires wet lab facilities, barcoded LNP formulations, and in vivo animal tumor models
Translational Design of Lipid Nanoparticles (LNPs) to Deliver mRNA Therapies to Solid Tumors and to the Lungs · Georgia Tech
Left open
Formulate top five ubiquibody constructs as mRNA in lipid nanoparticles and evaluate beta-catenin degradation in vitro and in mouse models. Blocker: Requires wet lab facilities, LNP formulation equipment, and animal model testing facilities
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