A facile approach for crosslinker free nano self assembly of protein for anti-tumor drug delivery: factors' optimization, characterization and in vitro evaluation

Sajid Asghar; Jumah Masoud M Salmani; Waseem Hassan; Ying Xie; Fanfei Meng; Zhigui Su; Minjie Sun; Yanyu Xiao; Qineng Ping

doi:10.1016/j.ejps.2014.06.022

A facile approach for crosslinker free nano self assembly of protein for anti-tumor drug delivery: factors' optimization, characterization and in vitro evaluation

Eur J Pharm Sci. 2014 Oct 15:63:53-62. doi: 10.1016/j.ejps.2014.06.022. Epub 2014 Jul 6.

Authors

Sajid Asghar¹, Jumah Masoud M Salmani², Waseem Hassan³, Ying Xie², Fanfei Meng², Zhigui Su², Minjie Sun², Yanyu Xiao², Qineng Ping⁴

Affiliations

¹ Department of Pharmaceutics, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China; College of Pharmacy, Government College University Faisalabad, Faisalabad, Pakistan.
² Department of Pharmaceutics, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China.
³ National Center for Drug Screening and State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China.
⁴ Department of Pharmaceutics, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China. Electronic address: pingqn@cpu.edu.cn.

PMID: 25004412
DOI: 10.1016/j.ejps.2014.06.022

Abstract

We report crosslinker free self assembly of bovine serum albumin (BSA) and a hydrophobic payload paclitaxel (PTX), into nanoparticles by harnessing the temperature driven unfolding of protein. To systematically study the effects of various factors responsible for the key attributes of the nanoparticles, a Resolution IV design was used. 20 formulations were made with pH, temperature, time of heating before and after addition of drug, stirring rate, protein concentration, and protein to drug ratio selected as independent variables. Particle size, encapsulation efficiency, yield and zeta potential were the response variables. Perturbation and Pareto charts were used to single out the important factors, while, mathematical equations and 3D surface charts have been used to describe the relationship between dependent and independent variables. Nanoparticles with size of 188-482 nm were observed with a highly negative zeta potential of -39.5 to -21.9. Nanoparticles obtained had decent encapsulation efficiency (72.5-87.9%) with effective yield (80.0-93.8%). Validation of the mathematical models with 4 runs indicated the good prognostic ability of Resolution IV design. Spectroscopic studies suggested the non-covalent complexation between BSA and PTX as the possible mechanism of self assembly due to irreversible conformational changes in protein. Transmission Electron Microscopy (TEM) revealed spherical nanoparticles with a porous network of PTX-BSA. X-ray Diffraction (XRD) showed amorphous nature of nanoparticles. PTX release from the nanoparticles was found to be controlled release and followed Peppas-Sahlin model. In vitro cytotoxicity of PTX-BSA nanoparticles was comparable to that of Taxol after 48 h treatment. These findings suggest heat driven BSA self assembly as a viable approach to formulate cytotoxic drug carrying nanoparticles which could be efficiently used in anti-cancer therapy.

Keywords: Bovine serum albumin; Nanoparticles; Paclitaxel; Resolution IV design; Self assembly.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antineoplastic Agents / chemistry
Antineoplastic Agents / pharmacology*
Cattle
Cell Proliferation / drug effects
Cross-Linking Reagents / chemistry*
Dose-Response Relationship, Drug
Drug Carriers / chemistry*
Drug Screening Assays, Antitumor
Hep G2 Cells
Humans
Nanoparticles / chemistry*
Particle Size
Serum Albumin, Bovine / chemistry*
Structure-Activity Relationship
Surface Properties

Substances

Antineoplastic Agents
Cross-Linking Reagents
Drug Carriers
Serum Albumin, Bovine