Lipoplex-induced hemagglutination: Potential involvement in intravenous gene delivery

N. Eliyahu, N. Servel, A. J. Domb, Y. Barenholz

Research output: Contribution to journalArticlepeer-review

131 Scopus citations


We report a study aiming to characterize the interaction of blood and blood components with lipoplexes under conditions relevant to in vivo intravenous transfection. In this study we focus on the interaction of lipoplexes with red blood cells (RBC). It was found that no significant hemolysis occurred during several hours' incubation using lipoplex compositions and lipoplex/red blood cell ratios in the range commonly used for in vivo transfection. However, the interaction of RBC with lipoplexes resulted in massive agglutination, which occurs irrespective of the type of cationic lipid or helper lipid. Agglutination was also induced by polyplexes (such as dendrimer/DNA complexes) and lipoplexes in the presence of spermidine or protamine sulfate (the latter induced hemagglutination by itself). DSPE-PEG2000 inserted into the lipoplexes inhibits hemagglutination somewhat. In order to understand the effect of serum on the agglutination better, plasma was separated into its high molecular weight components (HMWC, >14 kDa) and its low molecular weight components (LMWC, ≤14 kDa). These fractions were characterized for their level of proteins, primary amino groups, osmotic pressure, and electrical conductivity, and compared with saline (0.15 M NaCl). It was found that both LMWC and HMWC inhibit agglutination by themselves, although whole serum demonstrates better hemagglutination inhibition than each fraction separately. The inhibitory effect of the serum (or plasma) is explained by its effect on the electrostatics of the lipoplexes, reducing their positive charge, as was demonstrated using fluorescein-phosphatidylethanolamine-labeled lipoplexes. The effect of LMWC was related to ionic strength and was equal to the effect of 0.15 M NaCl. The level of agglutination was reduced with increasing lipoplex DNA/cationic lipid+ (DNA/L+) ratio. However, at the low DNA/L+ ratio needed to achieve significant in vivo transfection after i.v. administration, massive agglutination occurred. These data suggest that i.v. administration of lipoplexes and polyplexes may lead to RBC agglutination, and the agglutinates formed may explain the localization of lipoplexes and expression of their transgenes in the lungs.

Original languageEnglish
Pages (from-to)850-858
Number of pages9
JournalGene Therapy
Issue number13
StatePublished - 2002
Externally publishedYes

Bibliographical note

Funding Information:
This study was supported in part by grants from the Israel Science Foundation (ISF 8004/98) and the Canada– Israel Industrial Research and Development Foundation (CIIRDF).


  • Cationic lipids
  • Serum components
  • Transfection


Dive into the research topics of 'Lipoplex-induced hemagglutination: Potential involvement in intravenous gene delivery'. Together they form a unique fingerprint.

Cite this