Doxorubicin (Dox) was purchased from LC Laboratories (Woburn, MA) and functionalized with NHS-TCO-NHS linker as described below. Dox and Dox-TCO-NHS were resuspended in phosphate buffer solution (PBS, 100 mM sodium phosphate, 150 mM NaCl, pH 7.2, working buffer) for the bioconjugation studies. Tetrazine was synthesized according to previous reports.[4c (link),14 (link)] The nanoparticles (NPs) with an average particle size of 32 nm were synthesized according to our previous reports[15 (link)] and the brief procedure described below. The modified NPs were purified using Sephadex PD-10 columns purchased from GE Healthcare (Troy, NY). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from Corning Life Sciences (CellGro, Corning, NY) and supplemented with fetal bovine serum (FBS, Gibco, Thermo Fischer Scientific, Waltham, MA) and streptomycin/penicillin (Life Tech Corp., Grand Island, NY). Cy5.5-mono NHS ester was purchased from GE Healthcare and attached to the nanoparticles as described below.
Protocol detail
Doxorubicin-Tetrazine Nanoparticle Conjugation
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Doxorubicin (Dox)
Sephadex PD-10 columns
Dulbecco’s modified Eagle’s medium (DMEM)
Cy5.5-mono NHS ester
Brief procedure
Buffer
Cy5 5
Doxorubicin
Eagle
Ester
Nacl
Penicillin
Phosphate
Sephadex
Sodium phosphate
Streptomycin
Corresponding Organization : University at Albany, State University of New York
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Variable analysis
independent variables
- Doxorubicin (Dox) functionalization with NHS-TCO-NHS linker
- Synthesis of nanoparticles (NPs) with an average particle size of 32 nm
- Modification of NPs
- Attachment of Cy5.5-mono NHS ester to the nanoparticles
- None explicitly mentioned
- Phosphate buffer solution (PBS, 100 mM sodium phosphate, 150 mM NaCl, pH 7.2) used as working buffer for bioconjugation studies
- Tetrazine synthesized according to previous reports
- Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum (FBS) and streptomycin/penicillin
- None explicitly mentioned
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