Electrosomes, a novel carrier mediated drug delivery, targetted Drug delivery system, molecular pharmaceutics

1. 03/10/2025 1
MOLECULAR PHARMACEUTICS
ELECTROSOM
ES
GOKULRAJ S
M PHARM 2ND
SEMESTER
DEPARTMENT OF PHARMACEUTICS

2. 03/10/2025 2
MOLECULAR PHARMACEUTICS
INTRODUCTION
STRUCTURE OF ELECTROSOME
PREPARATION OF ELECTROSOMES
METHODS OF PREPARATION &
CHARACTERIZATION
ADVANTAGES & DISADVANTAGES
APPLICATIONS

3. 03/10/2025 3
MOLECULAR PHARMACEUTICS
• Electrosomes are a new and exciting area of research that has
potential application of pharmaceutical research and industry.
• Electrosomes are therapeutic system encapsulates therapeutics
agents within vesicles composed of electrically charged lipids or
polymers.
• Electrosomes are self assembled structures made up of
amphiphilic molecules, which can be charged by applying an
electric field.
• They typically in the size range from 50 to 500 nm.

4. 03/10/2025 4
MOLECULAR PHARMACEUTICS
Electrosomes are used to deliver,
• Proteins
• Peptides
• Other biological molecules that are difficult to deliver in
traditional way
Insulin – in treatment of
diabetes

5. 03/10/2025 5
MOLECULAR PHARMACEUTICS
STRUCTURE OF
ELECTROSOMES

6. 03/10/2025 6
MOLECULAR PHARMACEUTICS
• Electrosomes are self assembles structures made up of
amphiphilic molecules.
Hydrophilic and Hydrophobic regions
• These regions allow them to self assemble into unique structure
in response to an applied electric field.
• Structure can vary depending upon the type of amphiphilic
molecules used and conditions under which they are assembled.

7. 03/10/2025 7
MOLECULAR PHARMACEUTICS
Hydrophilic portion – outer shell
Hydrophobic portion - inner core

8. 03/10/2025 8
MOLECULAR PHARMACEUTICS
• The amphiphilic molecules can also be modified with targeting
ligands like
antibodies
peptides
to direct the Electrosomes to specific cells or tissue.
• Ligands attached to the Hydrophilic head group of the
Amphiphilic molecules.
• The electric filed can be used to control the size, shape and

9. 03/10/2025 9
MOLECULAR PHARMACEUTICS

10. 03/10/2025 10
MOLECULAR PHARMACEUTICS
PREPARATION OF ELECTOSOMES

11. 03/10/2025 11
MOLECULAR PHARMACEUTICS
MATERIALS
1. LIPIDS – phosphatidylcholine, cholesterol and cationic lipids like
DOTAP
2. DNA/RNA OR OTHER CARGO – plasmid DNA, RNA, or other
molecules to be delivered
3. ORGANIC SOLVENTS – chloroform, methanol
4. BUFFER SOLUTIONS – Phosphate Buffers Saline (PBS), Tris - EDTA
buffer

12. 03/10/2025 12
MOLECULAR PHARMACEUTICS
Preparation of lipid film
Hydration of lipid film
Formation of small Unilamellar vesicles
Incorporation of
DNA/RNA
Electroporation
Post
Electroporation
handling
Purification and
Characterization
ELECTROSOME

13. 03/10/2025 13
MOLECULAR PHARMACEUTICS
PREPARATION OF LIPID FILMS:
• Dissolve the lipids in a suitable organic solvents like chloroform
or Chloroform: Methanol
• Transfer the lipid solution to a round bottom flask
• Evaporate the solvent under reduced pressure using a rotary
evaporator to form a thin lipid film in the flask wall.
• Dry the lipid film under vacuum for several hours to remove
residual solvent.

14. 03/10/2025 14
MOLECULAR PHARMACEUTICS
HYDRATION OF LIPID FILM:
• Hydrate the lipid film with an aqueous buffer( eg: PBS or TE
buffer) by adding the buffer to the flask
• Vortex the mixture vigorously to form multilamellar vesicles.
FORMATION OF SUVs:
• Sonicate the MLV suspension using a probe sonicator until the
solution becomes clear, indicates the formation of SUVs.

15. 03/10/2025 15
MOLECULAR PHARMACEUTICS
INCORPORATION OF DNA/RNA:
• Mix the SUVs with the DNA/RNA solution.
• Allow the mixture to incubate for a specific time to enable the
DNA/RNA to interact with the liposomes.
ELECTROPORATION:
• Transfer the liposome-DNA/RNA mixture into an electroporation
cuvette.

16. 03/10/2025 16
MOLECULAR PHARMACEUTICS
• Set the electroporator to the desired voltage and capacitance
settings optimized for your liposome size and cargo.
• Apply the electric pulse to the mixture to facilitate the
encapsulation of DNA/RNA into the liposomes.
POST ELECTROPORATION HANDLING:
• Remove the mixture from the cuvette and transfer it to a fresh
tube. Allow the Electrosomes to recover for a specified period at
room temperature or 4ºC.

17. 03/10/2025 17
MOLECULAR PHARMACEUTICS
PURIFICATION:
• Optional
• Purify the Electrosomes to remove free DNA/RNA and
unencapsulated liposomes using techniques like
ultracentrifugation, size-exclusion chromatography, or dialysis.
CHARECTERIZATION:
• Characterization the Electrosomes for size, charge,
encapsulation efficiency and stability using techniques like
dynamic light scattering, zeta potential measurements and gel
electrophoresis.

18. 03/10/2025 18
MOLECULAR PHARMACEUTICS
METHODS OF PREPARATION

19. 03/10/2025 19
MOLECULAR PHARMACEUTICS
 Stains and constructs method
 Enzyme binding to Scaffolding
 Biofuel cell Assembly and Characterization
 Protein expression
 Enzyme active assays
 Construction of YSD of chimeric scaffolding
 Cyclic volumetric and Chronoamperometry

20. 03/10/2025 20
MOLECULAR PHARMACEUTICS
STRAIN AND CONSTRUCTS METHOD:
• Genes encoding dockerins of Acetivibrio cellulolyticus and
Clostrdium thermocellum were cloned and ligated to the C –
terminus of Zymomonad mobilis alcohol dehydrogenase and to
Pseudomonas putida formaldehyde dehydrogenase.
• The dockerin module of C. thermocellum was ligated to the C –
terminus of CueO of E.coli.

21. 03/10/2025 21
MOLECULAR PHARMACEUTICS
• All the dockerin containing enzymes encoding genes have been
cloned into the pET12b vector for expression in E>coli, yielding
the pET15b-zADH-AC, pFormDH-Ct and PET15b-CueO-Ct vectors.
• For control the gene encoding the native enzymes without an
appended dockerin module were also cloned in the same vector,
yielding plasmids pET15b-zADH, pET15b-pFormDH and pET15b-
CueO.

22. 03/10/2025 22
MOLECULAR PHARMACEUTICS
ENZYME BINDING TO SCAFFOLDIN:
• 2ml of yeast cells displaying Scaffoldin, were incubated with
bacterial lysates containing the expressed enzymes at room
temperature for 1hr.
• 1ml of bacterial lysates was diluted to 15ml and used for binding.
• As a binding buffer, 50mM Tris buffer at pH 8 with 1mM CaCl2 was
used after washing. Then the yeast cell were resuspended in 2ml
buffer for binding.

23. 03/10/2025 23
MOLECULAR PHARMACEUTICS
PROTIEN EXPRESSION:
• 10ml culture of E.coli was grown overnight at 37ºC.
• 1ml of culture was used to inoculate a 100ml culture in the same
medium containing Carbenicillin.
• Culture was incubated at 37ºC.
• 1mM Isopropyl ẞ-d-1-thiogalactopyranoside was added to
induce protein expression, followed by the overnight incubation
of culture at 20ºC

24. 03/10/2025 24
MOLECULAR PHARMACEUTICS
• The bacterial culture were lysed by sonication and lysate
containing the protein were separated by precipitation. The cells
were lysed in 50mM Tris buffer, pH 8 containing 1mM CaCl2.
• For cell expressing CueO, lysis were performed using 0.1M
acetate buffer, pH 5 containing 1mM CaCl2 and 800 microgram
CuSO4. For all lysate, bacterial culture were prepared in same
conditions in each culture to measure difference in enzyme
expression and activity level.

25. 03/10/2025 25
MOLECULAR PHARMACEUTICS
CHARACTERIZARION

26. 03/10/2025 26
MOLECULAR PHARMACEUTICS
PHYSICAL CHARECTERIZATION:
• Size and Size distribution
• Zeta potential (Surface potential)
ENCAPSULATION EFFICIENCY:
• Quantification of encapsulated drug by spectroscopy and
chromatography
STABILITY:
• Physical stability – leakage studies
- Size and Zeta potential over a period of time.

27. 03/10/2025 27
MOLECULAR PHARMACEUTICS
• Chemical stability – lipid peroxidation assay (measures oxidation of
lipids)
- pH stability test
BIOLOGIAL ACTIVITY:
• Cellular uptake flow – Cytometry (quantify the percentage of cells
that taken up Electrosomes using florescent labels)
- Confocal Microscopy – visualizes the
intracellular distribution of Electrosomes

28. 03/10/2025 28
MOLECULAR PHARMACEUTICS
• Gene Expression – reporter Gene assay
- RT qPCR (quantifies mRNA levels of delivered
gene)
• Cytotoxicity – MTT/MTS Assay (measures cell viability)
- Lactate Dehydrogenase Assay (Assesses cell
membrane integrity)
FUNCTIONAL ASSAY:
• In vitro & In vivo studies

29. 03/10/2025 29
MOLECULAR PHARMACEUTICS
APPLICATIONS

30. 03/10/2025 30
MOLECULAR PHARMACEUTICS
• They use enzymatic reactions to catalyze the conversion of
chemical energy to electricity in a fuel cell.
• The use of enzymatic cascades in enzymatic fuel cell anodes
resulted in very high-power outputs, as the electron density
achieved was much higher when the fuel was fully oxidized.
• It's used as a carrier in drug targeting.
• Used in the treatment of cancer.

31. 03/10/2025 31
MOLECULAR PHARMACEUTICS
• Used in studying immune response.
• Ear targeting Muscle targeting.
Some examples of drugs that have been used in Electrosomes
include
Lidocaine – local anesthetics
Fentanyl – pain relief
Rivastigmine – dementia treatment
Rotigotine – Parkinson’s disease treatment

32. 03/10/2025 32
MOLECULAR PHARMACEUTICS
ADVANTAGES
• Targeted drug delivery
• High drug loading capacity
• Precise control over size and shape - by varying the strength
and frequency of electric field.
• Stability - electric field makes them more resistant to
degradation and improved shelf life.

33. 03/10/2025 33
MOLECULAR PHARMACEUTICS
• Biocompatibility – lipids and polymers reduces the risk of toxicity
and immunogenicity.
• Non invasive administration
• Versatility – deliver wide range of proteins, peptides and nucleic
acid.
• Enhanced drug stability - hydrophobic core protect drugs from
degradation and oxidation.
• Improved bioavailability

34. 03/10/2025 34
MOLECULAR PHARMACEUTICS
• Controlled release
• Easy manufacturing
• Cost effective

35. 03/10/2025 35
MOLECULAR PHARMACEUTICS
DISADVANTAGES
• Complexity – requires specialized equipment and expertise
which increase the cost of production.
• Lack of FDA approval
• Limited scalability
• Immunogenicity – cationic lipids may stimulate the immune
response.
• Safety concern – long term safety is not assured; further

36. 03/10/2025 36
MOLECULAR PHARMACEUTICS
REFERENCES:
• Raktimava das sarkar, et al . Electrosomes as Novel
Nanocarrier. Indian Journal of Novel Drug Delivery,
2023:15(2);47-51.
• Kusuma Priya MD, et al. Somes: A review on composition,
formulation methods and evaluation of different types of
somes drug delivery system. International Journal of Applied
Pharmaceutics,2020:12(6);7-18.

37. 03/10/2025 37
MOLECULAR PHARMACEUTICS
• Szczupak A, Aizik D, Moraïs S, Vazana Y, Barak Y, Bayer A E,
Alfonta L. TheElectrosome: A Surface Displayed Enzymatic
Cascade in a Biofuel Cell's Anode and a High-Density Surface-
Displaye Biocathodic Enzyme. Nanomaterial.2017

38. 03/10/2025 38
MOLECULAR PHARMACEUTICS
THANKS FOR LISTENING