Spacer length and serum protein adsorption affect active targeting of trastuzumab-modified nanoparticles

Area of Science:

• Nanotechnology
• Biomedical Engineering
• Oncology

Background:

• Receptor-mediated active targeting of nanocarriers aims to improve cancer treatment specificity and reduce side effects.
• A major challenge is the adsorption of serum proteins, forming a 'protein corona' on nanocarriers in biological fluids.
• Understanding nanoparticle characteristics that influence protein corona formation is key for effective active targeting.

Purpose of the Study:

• To investigate the impact of different molecular weight spacers on the physicochemical properties of trastuzumab-conjugated human serum albumin nanoparticles.
• To evaluate the in vitro cell association of these nanoparticles with HER2-overexpressing (SK-BR-3) and control (MCF-7) cancer cells, with and without fetal bovine serum.
• To characterize the protein corona composition and assess its influence on the active targeting capability of the nanoparticles.

Main Methods:

• Covalent attachment of trastuzumab to human serum albumin nanoparticles using varying molecular weight spacers.
• Physicochemical characterization of the resulting nanoparticle formulations.
• In vitro cell association studies using SK-BR-3 and MCF-7 cell lines in the presence and absence of fetal bovine serum.
• Protein corona analysis using SDS-PAGE and LC-MS/MS.
• Evaluation of the influence of protein adsorption on active targeting.

Main Results:

• Different molecular weight spacers influenced nanoparticle physicochemical characteristics.
• Protein corona formation was observed, and its composition varied depending on nanoparticle formulation.
• Cell association studies demonstrated differences in targeting efficacy, influenced by the protein corona and HER2 expression.
• Trastuzumab-conjugated nanoparticles showed specific binding to HER2-overexpressing cells, modulated by protein adsorption.

Conclusions:

• Nanoparticle characteristics, including spacer length, significantly impact protein corona formation and subsequent cellular interactions.
• The protein corona plays a critical role in modulating the active targeting efficiency of nanocarriers in a biological environment.
• These findings provide insights for designing optimized nanocarrier systems for targeted cancer therapy, considering protein adsorption effects.