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1.
MAbs ; 10(2): 222-243, 2018.
Article in English | MEDLINE | ID: mdl-29293399

ABSTRACT

Antibody-drug conjugates (ADCs) are a growing class of biotherapeutics in which a potent small molecule is linked to an antibody. ADCs are highly complex and structurally heterogeneous, typically containing numerous product-related species. One of the most impactful steps in ADC development is the identification of critical quality attributes to determine product characteristics that may affect safety and efficacy. However, due to the additional complexity of ADCs relative to the parent antibodies, establishing a solid understanding of the major quality attributes and determining their criticality are a major undertaking in ADC development. Here, we review the development challenges, especially for reliable detection of quality attributes, citing literature and new data from our laboratories, highlight recent improvements in major analytical techniques for ADC characterization and control, and discuss newer techniques, such as two-dimensional liquid chromatography, that have potential to be included in analytical control strategies.


Subject(s)
Immunoconjugates/chemistry , Humans
2.
Int J Biochem Cell Biol ; 57: 54-62, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25310905

ABSTRACT

Recent studies have suggested that the receptor for advanced glycation end products (RAGE) participates in melanoma progression by promoting tumor growth. However, the mechanisms of RAGE activation in melanoma tumors are not clearly understood. To get deeper insights into these mechanisms, we transfected a melanoma cell line, which was established from a human melanoma primary tumor, with RAGE, and studied the effect of RAGE overexpression on cell proliferation and migration in vitro. We observed that overexpression of RAGE in these cells not only resulted in significantly increased migration rates compared to control cells, but also in decreased proliferation rates (Meghnani et al., 2014). In the present study, we compared the growth of xenograft tumors established from RAGE overexpressing WM115 cells, to that of control cells. We observed that when implanted in mice, RAGE overexpressing cells generated tumors faster than control cells. Analysis of protein tumor extracts showed increased levels of the RAGE ligands S100B, S100A2, S100A4, S100A6 and S100A10 in RAGE overexpressing tumors compared to control tumors. We show that the tumor growth was significantly reduced when the mice were treated with anti-RAGE antibodies, suggesting that RAGE, and probably several S100 proteins, were involved in tumor growth. We further demonstrate that the anti-RAGE antibody treatment significantly enhanced the efficacy of the alkylating drug dacarbazine in reducing the growth rate of RAGE overexpressing tumors.


Subject(s)
Melanoma/metabolism , Receptors, Immunologic/metabolism , S100 Proteins/biosynthesis , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/pharmacology , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/physiology , Dacarbazine/pharmacology , Female , Humans , Ligands , Melanoma/drug therapy , Mice , Mice, SCID , Random Allocation , Receptor for Advanced Glycation End Products , Receptors, Immunologic/antagonists & inhibitors , Receptors, Immunologic/immunology , Xenograft Model Antitumor Assays
3.
Methods Mol Biol ; 1045: 249-66, 2013.
Article in English | MEDLINE | ID: mdl-23913152

ABSTRACT

Antibodies are one of the most commonly used targeting ligands for nanocarriers, mainly because they are specific, have a strong binding affinity, and are available for a number of disease biomarkers. The bioconjugation chemistry can be a crucial factor in determining the targeting efficiency of drug delivery and should be chosen on a case-by-case basis. An antibody consists of a number of functional groups which offer many flexible options for bioconjugation. This chapter focuses on discussing some of the approaches including periodate oxidation, carbodiimide, maleimide, and heterofunctional linkers, for conjugating antibodies to different nanocarriers. The advantages and limitations are described herein. Specific examples are selected to demonstrate the experimental procedures and to illustrate the potential for applying to other nanocarrier system.


Subject(s)
Antibodies/chemistry , Drug Carriers/chemistry , Nanostructures/chemistry , Amines/chemistry , Carbodiimides/chemistry , Carbohydrates/chemistry , Carboxylic Acids/chemistry , Hydrazines/chemistry , Immunoglobulin G/chemistry , Oxidation-Reduction , Periodic Acid/chemistry , Succinimides/chemistry , Sulfhydryl Compounds/chemistry
4.
Small ; 9(12): 2129-39, 2013 Jun 24.
Article in English | MEDLINE | ID: mdl-23359548

ABSTRACT

The ability to map multiple biomarkers at the same time has far-reaching biomedical and diagnostic applications. Here, a series of biocompatible poly(D,L-lactic-co-glycolic acid) and polyethylene glycol particles for multicolor and multiplexed imaging are reported. More than 30 particle formulations that exhibit distinct emission signatures (ranging from the visible to NIR wavelength region) are designed and synthesized. These particles are encapsulated with combinations of carbocyanine-based fluorophores DiO, Dil, DiD, and DiR, and are characterized as <100 nm in size and brighter than commercial quantum dots. A particle formulation is identified that simultaneously emits fluorescence at three different wavelengths upon a single excitation at 485 nm via sequential and multiple FRET cascade events for multicolor imaging. Three other particles that display maximum fluorescence intensities at 570, 672, or 777 nm for multiplexed imaging are also identified. These particles are individually conjugated with specific (Herceptin or IgG2A11 antibody) or nonspecific (heptaarginine) ligands for targeting and, thus, could be applied to differentiate different cancer cells from a cell mixture according to the expressions of cell-surface human epidermal growth factor receptor 2 and the receptor for advanced glycation endproducts. Using an animal model subcutaneously implanted with the particles, it is further demonstrated that the developed platform could be useful for in vivo multiplexed imaging.


Subject(s)
Diagnostic Imaging/methods , Fluorescence Resonance Energy Transfer/methods , Nanoparticles/chemistry , Polymers/chemistry , Lactic Acid/chemistry , Polyethylene Glycols/chemistry , Polyglycolic Acid/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer
5.
Nanomedicine ; 9(4): 449-57, 2013 May.
Article in English | MEDLINE | ID: mdl-23178287

ABSTRACT

The cellular interactions and in vivo distribution of the nanomaterials are known to be strongly influenced by their physiochemical properties. Here, we investigated and compared the biocompatibility, pharmacokinetics, and biodistribution of previously reported peptide-based nanofiber (NFP), with commercially available nanomaterials. The NFP was a 2-dimensional (2D) structure with an extremely narrow width (4 nm) and a controllable length (50 to 400 nm). NFP was found to be non-toxic, hemocompatible, and with a minimum uptake by macrophages. In vivo studies further demonstrated that NFP could be delivered to the tumor site more effectively, and within a very shorter period of time, than spherical nanoparticles. Importantly, the undelivered NFP was rapidly eliminated by renal clearance and, thus, avoiding its accumulation in the spleen or liver. Overall, our data suggested a new paradigm in drug delivery via using a short circulating NFP, rather than a long circulating 3D nanoparticle, as a delivery cargo. FROM THE CLINICAL EDITOR: In this study, the role of small peptide-based nanocarriers is investigated for tumor-specific delivery, reporting excellent targeting properties and a favorable toxicity profile.


Subject(s)
Antineoplastic Agents/administration & dosage , Nanofibers , Neoplasms/drug therapy , Peptides/administration & dosage , Animals , Antineoplastic Agents/pharmacokinetics , Cell Line , Cell Line, Tumor , Humans , Mice , Mice, SCID , Tissue Distribution
6.
Analyst ; 137(23): 5487-90, 2012 Dec 07.
Article in English | MEDLINE | ID: mdl-23061092

ABSTRACT

Herein, we report the application of synthesized fluorescent, water soluble polymers for post-translational subtyping and differentiation of breast cancer cells in vitro. The fluorescence emission spectra from these polymers were modulated differently in the presence of conditioned cell culture media from various breast cancer cells. These polymers differentiate at a post-translation level possibly due to their ability to interact with extracellular enzymes that are over-expressed in cancerous conditions.


Subject(s)
Breast Neoplasms/classification , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Cell Line, Tumor , Culture Media, Conditioned , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Female , Fluorescent Dyes , HEK293 Cells , HeLa Cells , Humans , Metalloproteins/analysis , Metalloproteins/antagonists & inhibitors , Polymers , Spectrometry, Fluorescence
7.
ACS Nano ; 6(6): 4854-63, 2012 Jun 26.
Article in English | MEDLINE | ID: mdl-22559229

ABSTRACT

Semiconductor nanoparticles conjugated with organic- and dye-molecules to yield high efficiency visible photoluminescence (PL) hold great potential for many future technological applications. We show that folic acid (FA)-conjugated to nanosize TiO(2) and CeO(2) particles demonstrates a dramatic increase of photoemission intensity at wavelengths between 500 and 700 nm when derivatized using aminopropyl trimethoxysilane (APTMS) as spacer-linker molecules between the metal oxide and FA. Using density-functional theory (DFT) and time-dependent DFT calculations we demonstrate that the strong increase of the PL can be explained by electronic transitions between the titania surface oxygen vacancy (OV) states and the low-energy excited states of the FA/APTMS molecule anchored onto the surface oxygen bridge sites in close proximity to the OVs. We suggest this scenario to be a universal feature for a wide class of metal oxide nanoparticles, including nanoceria, possessing a similar band gap (∼3 eV) and with a large surface-vacancy-related density of electronic states. We demonstrate that the molecule-nanoparticle linker can play a crucial role in tuning the electronic and optical properties of nanosystems by bringing optically active parts of the molecule and of the surface close to each other.


Subject(s)
Metal Nanoparticles/chemistry , Metal Nanoparticles/ultrastructure , Organic Chemicals/chemistry , Oxides/chemistry , Crystallization/methods , Luminescence , Materials Testing , Particle Size , Surface Properties
8.
Bioconjug Chem ; 23(5): 981-92, 2012 May 16.
Article in English | MEDLINE | ID: mdl-22482883

ABSTRACT

The majority of near-infrared (NIR) fluorophores are organic molecules that show significant overlap between the excitation and emission spectra and therefore exhibit high fluorescence backgrounds during in vivo imaging. Recently, cyanine dyes with a large Stokes shift have shown great promise for NIR imaging but often undergo rapid photodegradation and nonspecific protein adsorption. Alternatively, fluorescence resonance energy transfer (FRET) is a promising technique to generate a larger gap between the excitation and emission maxima and thus can reduce the background signal. Here, we report the rational design of FRET-based polymeric nanoparticles for NIR and FRET imaging. The particles were assembled from diblock copolymers of poly(d,l-lactic-co-glycolic acid) and maleimide-activated poly(ethylene glycol), which were also encapsulated with both the donor (1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine) and acceptor (1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine) fluorophores. Because of their extreme hydrophobicity, thousands of fluorophores could be encapsulated inside a single particle without causing leakage. FRET resulted in a large Stokes shift (>100 nm) of the emission maxima, and the transfer efficiency could be fine-tuned by further adjusting the doping ratio of the donor and acceptor fluorophores. The optimized formulation was less than 100 nm in size, brighter than quantum dots, stable in biological media, and demonstrated similar biodistribution to most nanomaterials. Additional animal phantom studies demonstrated that the FRET imaging platform developed could have far-reaching applications in optical imaging.


Subject(s)
Carbocyanines/administration & dosage , Fluorescence Resonance Energy Transfer/methods , Fluorescent Dyes/administration & dosage , Nanoparticles/chemistry , Optical Imaging/methods , Animals , Carbocyanines/pharmacokinetics , Female , Fluorescent Dyes/pharmacokinetics , Hydrophobic and Hydrophilic Interactions , Lactic Acid/chemistry , Mice, SCID , Polyethylene Glycols/chemistry , Polyglycolic Acid/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer , Tissue Distribution
9.
Chem Commun (Camb) ; 46(18): 3209-11, 2010 May 14.
Article in English | MEDLINE | ID: mdl-20424776

ABSTRACT

A liposome-based amplified detection system is presented for the cancer cell secreted pathogenic enzyme matrix metalloproteinase-9 which does not require the use of biological antibodies.


Subject(s)
Liposomes/chemistry , Matrix Metalloproteinase 9/analysis , Phenylenediamines/metabolism , Cell Line, Tumor , Horseradish Peroxidase/metabolism , Humans
10.
Bioconjug Chem ; 20(7): 1332-9, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19601658

ABSTRACT

Liposomes have been widely used as a drug delivery vehicle, and currently, more than 10 liposomal formulations are approved by the Food and Drug Administration for clinical use. However, upon targeting, the release of the liposome-encapsulated contents is usually slow. We have recently demonstrated that contents from appropriately formulated liposomes can be rapidly released by the cancer-associated enzyme matrix metalloproteinase-9 (MMP-9). Herein, we report our detailed studies to optimize the liposomal formulations. By properly selecting the lipopeptide, the major lipid component, and their relative amounts, we demonstrate that the contents are rapidly released in the presence of cancer-associated levels of recombinant human MMP-9. We observed that the degree of lipid mismatch between the lipopepides and the major lipid component profoundly affects the release profiles from the liposomes. By utilizing the optimized liposomal formulations, we also demonstrate that cancer cells (HT-29) which secrete low levels of MMP-9 failed to release a significant amount of the liposomal contents. Metastatic cancer cells (MCF7) secreting high levels of the enzyme rapidly release the encapsulated contents from the liposomes.


Subject(s)
Liposomes/chemistry , Liposomes/metabolism , Matrix Metalloproteinase 9/metabolism , Adenocarcinoma/enzymology , Breast Neoplasms/enzymology , Cell Line, Tumor , Colorectal Neoplasms/enzymology , Female , Fluoresceins/analysis , Humans , Lipopeptides/chemical synthesis , Lipopeptides/chemistry
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