Biotin-Conjugated Polychelating Agent

Torchilin, Vladimir P.

doi:10.1021/bc9800567

Cited by 14 publications

(12 citation statements)

References 9 publications

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“…In order to evaluate the effects of charge on nuclear uptake, we carried out an additional set of experiments with a sample of tmFab-SAv-Bi-MCP that was saturated with stable In 3+ after being labeled with 111 In. We have previously shown that there is no loss of free 111 In 3+ ions under these conditions. − The MCP in tmFab-SAv-Bi-MCP carries approximately 50 DTPA pendant groups. When labeled with 111 In 3+ , only a small fraction of these groups bind In ions.…”

Section: Resultsmentioning

confidence: 90%

Intracellular Routing in Breast Cancer Cells of Streptavidin-Conjugated Trastuzumab Fab Fragments Linked to Biotinylated Doxorubicin-Functionalized Metal Chelating Polymers

et al. 2014

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We describe the synthesis of a heterotelechelic metal-chelating polymer (Bi-MCP-Dox), a polyacrylamide with a number average degree of polymerization DPn = 50 (PDI = 1.2), with biotin (Bi) and doxorubicin (Dox) as functional chain ends and diethylenetriaminepentaacetic acid (DTPA) pendant groups as the binding sites for metal ions. We compared its behavior in cell-uptake experiments with a similar polymer (Bi-MCP) without Dox. These MCPs were complexed with trastuzumab Fab (tmFab) fragments covalently linked to streptavidin (SAv) to form tmFab-SAv-Bi-MCP-Dox and tmFab-SAv-Bi-MCP via the strong affinity between Bi and SAv. tmFab targets human epidermal growth factor receptor-2 (HER2), which is overexpressed on certain human breast cancer cells. Surface plasmon resonance (SPR) experiments with the extracellular domain (ECD) of HER2 showed that incorporation of the MCPs in these complexes had no significant effect on the association or dissociation rate with the HER2 ECD and the dissociation constants. The tmFab-complexed MCPs were subsequently labeled with (111)In (an Auger electron emitting radionuclide). Auger electrons can cause lethal DNA double strand breaks (DSBs) but only if they are emitted intracellularly and especially, in close proximity to the nucleus. To evaluate the cellular and nuclear uptake of tmFab-SAv-Bi-MCP-Dox, we incubated HER2+ SK-BR-3 human breast cancer cells with the complexes saturated with stable In(3+) and visualized their distribution by confocal fluorescence microscopy, monitoring the fluorescence of Dox. In parallel, we carried out cell fractionation studies on tmFab-SAv-Bi-MCP-Dox and on tmFab-SAv-Bi-MCP labeled with (111)In. Both radiolabeled complexes showed cell internalization and nuclear localization. We conclude that metal-chelating polymers with this composition appear to encourage internalization, nuclear uptake, and chromatin (DNA) binding of trastuzumab fragments modified with streptavidin in human breast cancer cells expressing HER2. Further study is needed to understand the impact of polymer charge on cellular uptake and distribution to intracellular compartments.

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Section: Resultsmentioning

confidence: 90%

Intracellular Routing in Breast Cancer Cells of Streptavidin-Conjugated Trastuzumab Fab Fragments Linked to Biotinylated Doxorubicin-Functionalized Metal Chelating Polymers

et al. 2014

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“…As biotin is a small molecule with few functions, it is not straightforward. Nevertheless, some authors suggested use of poly-L-lysine (PL) to covalently bind both biotin and the reporter groups with a chosen ratio (19,20). Other hapten tracers have been prepared in this manner (21)(22)(23).…”

Section: Resultsmentioning

confidence: 99%

Transition Metal−Carbonyl Labeling of Biotin and Avidin for Use in Solid-Phase Carbonyl Metallo Immunoassay (CMIA)

et al. 2002

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The preparation of several transition metal-carbonyl tracers of biotin and avidin is described. Multiple labeling of avidin was achieved by acylation of some of its amine-bearing residues with N-succinimidyl 4-pentynoate (dicobalt hexacarbonyl). By varying the initial amount of this complex, protein conjugates with the extent of derivatization of up to 13 were obtained. Biotin was labeled with one (eta5-cyclopentadienyl)manganese tricarbonyl moiety by reaction of biotin hydrazide, whereas multiple labeling was reached by successive conjugation of biotin and N-succinimidyl 4-pentynoate (dicobalt hexacarbonyl) or (eta5-cyclopentadienyl)iron dicarbonyl (eta1-N-maleimidato) to poly-L-lysine or fourth generation Starburst dendrimer. All the conjugates displayed a good to excellent bioaffinity toward their respective counterparts, as measured by competitive enzymatic assays.

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“…Applications utilizing polylysine as carriers for nuclides have previously been proposed, such as a polychelating agent for high-specific-activity metal-nuclide binding of antibodies, for use in single-photon emission computed tomography (SPECT) and magnetic resonance tomography, and for therapy (29). The application of polylysine as an effector carrier for use in PRIT has also been reported (17,18).…”

Section: Discussionmentioning

confidence: 99%

“…In a few investigations, poly- l -lysine has been explored as a multicarrier of biotin and radionuclides for pretargeted radioimmunotherapy ( , ). The advantages of these polymer conjugates are that increased avidity for avidin and higher specific radioactivity may be achieved.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and Biodistribution of ²¹¹At-Labeled, Biotinylated, and Charge-Modified Poly-l-lysine: Evaluation for Use as an Effector Molecule in Pretargeted Intraperitoneal Tumor Therapy

et al. 2002

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Poly-L-lysine (7, 21, and 204 kDa) has been evaluated as an effector carrier for use in pretargeted intraperitoneal tumor therapy. For the synthesis, the epsilon-amino groups on the poly-L-lysine were modified in three steps utilizing conjugate biotinylation with biotin amidocaproate N-hydroxysuccinimide ester (BANHS), conjugate radiolabeling with (211)At using the intermediate reagent N-succinimidyl 3-(trimethylstannyl)benzoate (m-MeATE), and charge modification using succinic anhydride, resulting in an increase in the molecular weight of approximately 80% of the final product. The labeling of the m-MeATE reagent and subsequent conjugation of the polymer were highly efficient with overall radiochemical yields in the range of 60-70%. The in vitro avidin binding ability of the modified polymer was almost complete (90-95%), as determined by binding to avidin beads using a convenient filter tube assay. Following intraperitoneal (ip) injection in athymic mice, the 13 kDa polymer product was cleared mainly via the kidneys with fast kinetics (biological half-live T(b) approximately 2 h) and with low whole-body retention. The clearance of the 38 kDa polymer was distributed between kidneys and liver, and the 363 kDa polymer was mainly sequestered by the liver with a T(b) of 8 h. Increased tissue uptake in the thyroid, lungs, stomach, and spleen following the distribution of the large effector molecules (38 and 363 kDa) suggests that degradation of the polymers by the liver may release some of the label as free astatine/astatide.

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Biotin-Conjugated Polychelating Agent

Cited by 14 publications

References 9 publications

Intracellular Routing in Breast Cancer Cells of Streptavidin-Conjugated Trastuzumab Fab Fragments Linked to Biotinylated Doxorubicin-Functionalized Metal Chelating Polymers

Intracellular Routing in Breast Cancer Cells of Streptavidin-Conjugated Trastuzumab Fab Fragments Linked to Biotinylated Doxorubicin-Functionalized Metal Chelating Polymers

Transition Metal−Carbonyl Labeling of Biotin and Avidin for Use in Solid-Phase Carbonyl Metallo Immunoassay (CMIA)

Synthesis and Biodistribution of ²¹¹At-Labeled, Biotinylated, and Charge-Modified Poly-l-lysine: Evaluation for Use as an Effector Molecule in Pretargeted Intraperitoneal Tumor Therapy

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Biotin-Conjugated Polychelating Agent

Cited by 14 publications

References 9 publications

Intracellular Routing in Breast Cancer Cells of Streptavidin-Conjugated Trastuzumab Fab Fragments Linked to Biotinylated Doxorubicin-Functionalized Metal Chelating Polymers

Intracellular Routing in Breast Cancer Cells of Streptavidin-Conjugated Trastuzumab Fab Fragments Linked to Biotinylated Doxorubicin-Functionalized Metal Chelating Polymers

Transition Metal−Carbonyl Labeling of Biotin and Avidin for Use in Solid-Phase Carbonyl Metallo Immunoassay (CMIA)

Synthesis and Biodistribution of 211At-Labeled, Biotinylated, and Charge-Modified Poly-l-lysine: Evaluation for Use as an Effector Molecule in Pretargeted Intraperitoneal Tumor Therapy

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Synthesis and Biodistribution of ²¹¹At-Labeled, Biotinylated, and Charge-Modified Poly-l-lysine: Evaluation for Use as an Effector Molecule in Pretargeted Intraperitoneal Tumor Therapy