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Microfluidic Synthesis of PEG- and Folate-Conjugated Liposomes for One-Step Formation of Targeted Stealth Nanocarriers

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ABSTRACT

Purpose

A microfluidic hydrodynamic flow focusing technique enabling the formation of small and nearly monodisperse liposomes is investigated for continuous-flow synthesis of poly(ethylene glycol) (PEG)-modified and PEG-folate-functionalized liposomes for targeted drug delivery.

Methods

Controlled laminar flow in thermoplastic microfluidic devices facilitated liposome self-assembly from initial lipid compositions including lipid/cholesterol mixtures containing PEG-lipid and folate-PEG-lipid conjugates. Relationships among flow conditions, lipid composition, and liposome size were evaluated; their impact on PEG and folate incorporation were determined through a combination of UV–vis absorbance measurements and characterization of liposome zeta potential.

Results

PEG and folate were successfully incorporated into microfluidic-synthesized liposomes over the full range of liposome sizes studied. Efficiency of PEG-lipid incorporation was inversely correlated with liposome diameter. Folate-lipid was effectively integrated into liposomes at various flow conditions.

Conclusions

Liposomes incorporating relatively large PEG-modified and folate-PEG-modified lipids were successfully synthesized using the microfluidic flow focusing platform, providing a simple, low cost, rapid method for preparing functionalized liposomes. Relationships between preparation conditions and PEG or folate-PEG functionalization have been elucidated, providing insight into the process and defining paths for optimization of the microfluidic method toward the formation of functionalized liposomes for pharmaceutical applications.

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Abbreviations

AF4 :

asymmetric flow field-flow fractionation

Cryo-TEM:

cryogenic temperature transmission electron microscopy

DCP:

dihexadecyl phosphate

DMPC:

1,2-dimyristoyl-sn-glycero-3-phosphocholine

DSPE-PEG2000-folate:

1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[folate(PEG)-2000]

FPL:

folate–PEG-modified liposomes

FR:

folate receptor

FRR:

flow rate ratio

FWHM:

full width at half maximum

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

MALLS:

multiangle laser light scattering

PEG:

poly(ethylene glycol)

PEG2000-PE:

1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(PEG)-2000]

PEG5000-PE:

1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(PEG)-5000

PL:

PEG-modified (PEGylated)

QELS:

quasi-elastic light scattering

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ACKNOWLEDGMENTS AND DISCLOSURES

Cryo-TEM imaging was performed at the Nanoscale Imaging, Spectroscopy, and Properties (NISP) Laboratory of the Maryland NanoCenter at the University of Maryland, College Park. This research was supported by NIH grants R21EB011750 and R21EB009485, NSF grant CBET0966407, NIST-ARRA Fellowship Program administered by the University of Maryland, and the NRC Research Associateship Program.

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Authors and Affiliations

  1. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland, 20742, USA

    Renee R. Hood & Don L. DeVoe

  2. Department of Mechanical Engineering, University of Maryland, College Park, Maryland, 20742, USA

    Chenren Shao & Don L. DeVoe

  3. Biochemical Sciences Division, National Institute of Standards and Technology, Gaithersburg, Maryland, 20899-6313, USA

    Renee R. Hood, Donna M. Omiatek & Wyatt N. Vreeland

  4. University of Maryland, 3139 Glenn L. Martin Hall, Building 088, College Park, Maryland, 20742, USA

    Don L. DeVoe

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  1. Renee R. Hood
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Correspondence to Don L. DeVoe.

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Hood, R.R., Shao, C., Omiatek, D.M. et al. Microfluidic Synthesis of PEG- and Folate-Conjugated Liposomes for One-Step Formation of Targeted Stealth Nanocarriers. Pharm Res 30, 1597–1607 (2013). https://doi.org/10.1007/s11095-013-0998-3

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  • DOI: https://doi.org/10.1007/s11095-013-0998-3

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