Evaluation of interstitial protein delivery in multicellular layers model

Soo Yeon Kim, Tae Hyung Kim, Jong Hoon Choi, Kang Choon Lee, Ki Dong Park, Seung Jin Lee, Hyo Jeong Kuh

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


The limited efficacy of anticancer protein drugs is related to their poor distribution in tumor tissue. We examined interstitial delivery of four model proteins of different molecular size and bioaffinity in multicellular layers (MCL) of human cancer cells. Model proteins were tumor necrosis factor-related apoptosis-including ligand (TRAIL), cetuximab, RNase A, and IgG. MCLs were cultured in Transwell inserts, exposed to drugs, then cryo-sectioned for image acquisition using fluorescence microscopy (fluorescent dye-labeled TRAIL, RNase A, IgG) or immunohistochemistry (cetuximab). TRAIL and cetuximab showed partial penetration into MCLs, whereas RNase A and IgG showed insignificant penetration. At 10-fold higher dose, a significant increase in penetration was observed for IgG only, while cetuximab showed an intense accumulation limited to the front layers. PEGylated TRAIL and RNase A formulated in a heparin-Pluronic (HP) nanogel showed significantly improved penetration attributable to increased stability and extracellular matrix binding, respectively. IgG penetration was significantly enhanced with paclitaxel pretreatment as a penetration enhancer. The present study suggests that MCL culture may be useful in evaluation of protein delivery in the tumor interstitium. Four model proteins showed limited interstitial penetration in MCL cultures. Bioaffinity, rather than molecular size, seems to have a positive effect on tissue penetration, although high binding affinity may lead to sequestration in the front cell layers. Polymer conjugation and nanoformulation, such as PEGylation and HP nanogel, or use of penetration enhancers are potential strategies to increase interstitial delivery of anticancer protein drugs.

Original languageEnglish
Pages (from-to)531-541
Number of pages11
JournalArchives of Pharmacal Research
Issue number3
StatePublished - Mar 2012

Bibliographical note

Funding Information:
This work was supported by the Nano-Biotechnology Project (Regenomics, 2011-0007745) of the Korean Science and Engineering Foundation (KOSEF) and Mid-career researcher program through NRF grant funded by the MEST (No. 2011-0027565). S-Y Kim was supported by Korea Industrial Technology Foundation (KOTEF) through the Human Resource Trainig Project for Strategic Technology.


  • Cetuximab
  • Heparin-Pluronic nanogels
  • Multicellular layers
  • PEGylation
  • Protein delivery


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