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Antibody-mediated targeting of iron oxide nanoparticles to the folate receptor alpha increases tumor cell association in vitro and in vivo.

Ndong C, Toraya-Brown S, Kekalo K, Baker I, Gerngross TU, Fiering SN, Griswold KE - Int J Nanomedicine (2015)

Bottom Line: Here, we describe molecular targeting of iron oxide nanoparticles (IONPs) to the folate receptor alpha (FOLRα) using an engineered antibody fragment (Ffab).Interestingly, Ffab-IONPs built with custom 120 nm nanoparticles exhibited lower in vitro targeting efficiency when compared to those built with commercially sourced 180 nm nanoparticles.In vivo, however, the two Ffab-IONP platforms achieved equivalent tumor homing, although the smaller 120 nm IONPs were more prone to liver sequestration.

View Article: PubMed Central - PubMed

Affiliation: Thayer School of Engineering, Dartmouth, Hanover, NH, USA.

ABSTRACT
Active molecular targeting has become an important aspect of nanoparticle development for oncology indications. Here, we describe molecular targeting of iron oxide nanoparticles (IONPs) to the folate receptor alpha (FOLRα) using an engineered antibody fragment (Ffab). Compared to control nanoparticles targeting the non-relevant botulinum toxin, the Ffab-IONP constructs selectively accumulated on FOLRα-overexpressing cancer cells in vitro, where they exhibited the capacity to internalize into intracellular vesicles. Similarly, Ffab-IONPs homed to FOLRα-positive tumors upon intraperitoneal administration in an orthotopic murine xenograft model of ovarian cancer, whereas negative control particles showed no detectable tumor accumulation. Interestingly, Ffab-IONPs built with custom 120 nm nanoparticles exhibited lower in vitro targeting efficiency when compared to those built with commercially sourced 180 nm nanoparticles. In vivo, however, the two Ffab-IONP platforms achieved equivalent tumor homing, although the smaller 120 nm IONPs were more prone to liver sequestration. Overall, the results show that Ffab-mediated targeting of IONPs yields specific, high-level accumulation within cancer cells, and this fact suggests that Ffab-IONPs could have future utility in ovarian cancer diagnostics and therapy.

No MeSH data available.


Related in: MedlinePlus

Schematic of workflow for fab and IONP functionalization.Notes: Monomeric Ffab and Bfab are subjected to reduction/activation using 20 mM cysteine followed by conjugation with maleimiede-PEG2-biotin or maleimide-activated IONPs. Two IONP types were examined in this study: Dartmouth CMD and commercial BNF, which were functionalized with maleimide groups using EDC and N-(2-aminoethyl) maleimide, or sulfo-GMBS, respectively.Abbreviations: IONP, iron oxide nanoparticle; fab, an engineered monoclonal antibody fragment; Ffab, Farletuzufab, engineered from monoclonal antibody Farletuzumab; Bfab, anti-botulinum toxin fab fragment; PEG2, polyethylene glycol 2; CMD, carboxymethyl-dextran; BNF, bionized nanoferrite; EDC, 1-ethyl-3-(3-dimethylaminopropyl) carboiimide; sulfo-GMBS, N-(γ-maleimobutyryloxy) sulfosuccinimide ester.
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f1-ijn-10-2595: Schematic of workflow for fab and IONP functionalization.Notes: Monomeric Ffab and Bfab are subjected to reduction/activation using 20 mM cysteine followed by conjugation with maleimiede-PEG2-biotin or maleimide-activated IONPs. Two IONP types were examined in this study: Dartmouth CMD and commercial BNF, which were functionalized with maleimide groups using EDC and N-(2-aminoethyl) maleimide, or sulfo-GMBS, respectively.Abbreviations: IONP, iron oxide nanoparticle; fab, an engineered monoclonal antibody fragment; Ffab, Farletuzufab, engineered from monoclonal antibody Farletuzumab; Bfab, anti-botulinum toxin fab fragment; PEG2, polyethylene glycol 2; CMD, carboxymethyl-dextran; BNF, bionized nanoferrite; EDC, 1-ethyl-3-(3-dimethylaminopropyl) carboiimide; sulfo-GMBS, N-(γ-maleimobutyryloxy) sulfosuccinimide ester.

Mentions: The workflow for IONP functionalization and subsequent conjugation with the engineered antibodies is shown in Figure 1. The two antibody fragments, Farletuzufab (Ffab), targeting the FOLRα cancer antigen, and Botulifab (Bfab), targeting the negative control protein botulinum toxin, were constructed such that each retained an unpaired C-terminal cysteine on the heavy chain. This design ultimately enabled site-specific conjugation to either maleimide-PEG2-biotin-or maleimide-functionalized IONPs. Ffab and Bfab were expressed in HEK 293 cells, and the desired fab monomers were isolated by Kappa select affinity purification followed by size exclusion chromatography (Figure S1A, B).


Antibody-mediated targeting of iron oxide nanoparticles to the folate receptor alpha increases tumor cell association in vitro and in vivo.

Ndong C, Toraya-Brown S, Kekalo K, Baker I, Gerngross TU, Fiering SN, Griswold KE - Int J Nanomedicine (2015)

Schematic of workflow for fab and IONP functionalization.Notes: Monomeric Ffab and Bfab are subjected to reduction/activation using 20 mM cysteine followed by conjugation with maleimiede-PEG2-biotin or maleimide-activated IONPs. Two IONP types were examined in this study: Dartmouth CMD and commercial BNF, which were functionalized with maleimide groups using EDC and N-(2-aminoethyl) maleimide, or sulfo-GMBS, respectively.Abbreviations: IONP, iron oxide nanoparticle; fab, an engineered monoclonal antibody fragment; Ffab, Farletuzufab, engineered from monoclonal antibody Farletuzumab; Bfab, anti-botulinum toxin fab fragment; PEG2, polyethylene glycol 2; CMD, carboxymethyl-dextran; BNF, bionized nanoferrite; EDC, 1-ethyl-3-(3-dimethylaminopropyl) carboiimide; sulfo-GMBS, N-(γ-maleimobutyryloxy) sulfosuccinimide ester.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4388088&req=5

f1-ijn-10-2595: Schematic of workflow for fab and IONP functionalization.Notes: Monomeric Ffab and Bfab are subjected to reduction/activation using 20 mM cysteine followed by conjugation with maleimiede-PEG2-biotin or maleimide-activated IONPs. Two IONP types were examined in this study: Dartmouth CMD and commercial BNF, which were functionalized with maleimide groups using EDC and N-(2-aminoethyl) maleimide, or sulfo-GMBS, respectively.Abbreviations: IONP, iron oxide nanoparticle; fab, an engineered monoclonal antibody fragment; Ffab, Farletuzufab, engineered from monoclonal antibody Farletuzumab; Bfab, anti-botulinum toxin fab fragment; PEG2, polyethylene glycol 2; CMD, carboxymethyl-dextran; BNF, bionized nanoferrite; EDC, 1-ethyl-3-(3-dimethylaminopropyl) carboiimide; sulfo-GMBS, N-(γ-maleimobutyryloxy) sulfosuccinimide ester.
Mentions: The workflow for IONP functionalization and subsequent conjugation with the engineered antibodies is shown in Figure 1. The two antibody fragments, Farletuzufab (Ffab), targeting the FOLRα cancer antigen, and Botulifab (Bfab), targeting the negative control protein botulinum toxin, were constructed such that each retained an unpaired C-terminal cysteine on the heavy chain. This design ultimately enabled site-specific conjugation to either maleimide-PEG2-biotin-or maleimide-functionalized IONPs. Ffab and Bfab were expressed in HEK 293 cells, and the desired fab monomers were isolated by Kappa select affinity purification followed by size exclusion chromatography (Figure S1A, B).

Bottom Line: Here, we describe molecular targeting of iron oxide nanoparticles (IONPs) to the folate receptor alpha (FOLRα) using an engineered antibody fragment (Ffab).Interestingly, Ffab-IONPs built with custom 120 nm nanoparticles exhibited lower in vitro targeting efficiency when compared to those built with commercially sourced 180 nm nanoparticles.In vivo, however, the two Ffab-IONP platforms achieved equivalent tumor homing, although the smaller 120 nm IONPs were more prone to liver sequestration.

View Article: PubMed Central - PubMed

Affiliation: Thayer School of Engineering, Dartmouth, Hanover, NH, USA.

ABSTRACT
Active molecular targeting has become an important aspect of nanoparticle development for oncology indications. Here, we describe molecular targeting of iron oxide nanoparticles (IONPs) to the folate receptor alpha (FOLRα) using an engineered antibody fragment (Ffab). Compared to control nanoparticles targeting the non-relevant botulinum toxin, the Ffab-IONP constructs selectively accumulated on FOLRα-overexpressing cancer cells in vitro, where they exhibited the capacity to internalize into intracellular vesicles. Similarly, Ffab-IONPs homed to FOLRα-positive tumors upon intraperitoneal administration in an orthotopic murine xenograft model of ovarian cancer, whereas negative control particles showed no detectable tumor accumulation. Interestingly, Ffab-IONPs built with custom 120 nm nanoparticles exhibited lower in vitro targeting efficiency when compared to those built with commercially sourced 180 nm nanoparticles. In vivo, however, the two Ffab-IONP platforms achieved equivalent tumor homing, although the smaller 120 nm IONPs were more prone to liver sequestration. Overall, the results show that Ffab-mediated targeting of IONPs yields specific, high-level accumulation within cancer cells, and this fact suggests that Ffab-IONPs could have future utility in ovarian cancer diagnostics and therapy.

No MeSH data available.


Related in: MedlinePlus