title: Influence of molecular weight of PEG chain on interaction between streptavidin and biotin-PEG-conjugated phospholipids studied with QCM-D.
abstract: Poly(ethylene glycol)-conjugated phospholipid (PEG-lipid) derivatives spontaneously incorporate into lipid bilayer membranes, thus, they are useful for immobilizing bioactive substances onto cell surfaces. Here, we investigated how the density and molecular weight of PEG molecules influenced immobilization and cellular uptake of a bioactive substance. We analyzed how three biotin-PEG-lipids (1k, 5k, and 40k, with PEG molecular weights: 1kD, 5kD, and 40kD, respectively) interacted with streptavidin on a surface attached to a quartz crystal microbalance with dissipation (QCM-D). We found that the volume excluded by 1k PEG could not effectively prevent adsorption of bovine serum albumin (BSA). In contrast, 5k PEG chains could completely prevent protein adsorption. However, due to strong static repulsion, 40k PEG chains could not be packed at high density. Thus, BSA migrated between PEG chains, and adsorption was not effectively prevented. When streptavidin was added, it bound to multiple neighboring sites on 1k and 5k biotin-PEG-lipids, which reduced chain viscoelasticity. In contrast, streptavidins bound at a one-to-one stoichiometry with the 40k biotin-PEG-lipids, which increased chain viscoelasticity. However, differences in PEG viscoelasticity and PEG molecular weights did not influence cellular uptake of immobilized streptavidin. Therefore, these are not important factors in designing polymers that prevent cellular endocytosis.
  
  
  CONCLUSIONS
  Poly(ethylene glycol)-conjugated phospholipid (PEG-lipid) derivatives have been widely used to modify not only liposome surface, but also the surfaces of cells and pancreatic islets for cell transplantation. Since the entire cell surface can be modified with PEG-lipid through hydrophobic interactions, it is a promising approach for improving graft survival in clinical settings. Although the surface modification is protective in the early stages of transplantation, it is important to understand the factors that influence on the cellular uptake. In this study, we examined the influence of the surface density and molecular weights of PEG-lipids on the cellular uptake by QCM-D and cellular experiments. It was found that the differences in viscoelasticity of PEG chain did not affect on the cellular uptake.
authors.affiliation: Department of Bioengineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. Electronic address: teramura@bioeng.t.u-tokyo.ac.jp.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Teramura
authors.affiliation: Department of Bioengineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Kuroyama
authors.affiliation: Department of Bioengineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Madoka
authors.initials: M
authors.lastname: Takai
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry*; D002417:Cattle; D045744:Cell Line, Tumor; D006801:Humans; D008970:Molecular Weight; D010743:Phospholipids / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D058525:Quartz Crystal Microbalance Techniques / Q000379:methods; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D010743:Phospholipids; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D001710:Biotin; D019809:Streptavidin; C000595209:polyethylene glycol 1000
keywords: Biotin; Poly(ethylene glycol)-conjugated phospholipid (PEG-lipid); QCM-D; Streptavidin; Surface modification