A novel assay for monitoring internalization of nanocarrier coupled antibodies.

A novel assay for monitoring internalization of nanocarrier coupled antibodies. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

A novel assay for monitoring internalization of nanocarrier coupled antibodies. Abstract Text:

Ulrik B Nielsen,Dmitri B Kirpotin,Edward M Pickering,Daryl C Drummond,James D Marks,

BACKGROUND: Discovery of tumor-selective antibodies or antibody fragments is a promising approach for delivering therapeutic agents to antigen over-expressing cancers. Therefore it is important to develop methods for the identification of target- and function specific antibodies for effective drug delivery. Here we describe a highly selective and sensitive method for characterizing the internalizing potential of multivalently displayed antibodies or ligands conjugated to liposomes into tumor cells. The assay requires minute amounts of histidine-tagged ligand and relies on the non-covalent coupling of these antibodies to fluorescent liposomes containing a metal ion-chelating lipid. Following incubation of cells with antibody-conjugated liposomes, surface bound liposomes are gently removed and the remaining internalized liposomes are quantitated based on fluorescence in a high throughput manner. We have termed this methodology "Chelated Ligand Internalization Assay", or CLIA. RESULTS: The specificity of the assay was demonstrated with different antibodies to the ErbB-2 and EGF receptors. Antibody-uptake correlated with receptor expression levels in tumor cell lines with a range of receptor expression. Furthermore, Ni-NTA liposomes containing doxorubicin were used to screen for the ability of antibodies to confer target-specific cytotoxicity. Using an anti-ErbB2 single chain Fv (scFv) (F5) antibody, cytotoxicity could be conferred to ErbB2-overexpressing cells; however, a poly(ethylene glycol)-linked lipid (DSPE-PEG-NTA-Ni) was necessary to allow for efficient loading of the drug and to reduce nonspecific drug leakage during the course of the assay. CONCLUSION: The CLIA method we describe here represents a rapid, sensitive and robust assay for the identification and characterization of tumor-specific antibodies capable of high drug-delivery efficiency when conjugated to liposomal nanocarriers.

A novel assay for monitoring internalization of nanocarrier coupled antibodies. Publishing Authors By Initials

UB Nielsen,DB Kirpotin,EM Pickering,DC Drummond,JD Marks,

For similar investigative techniques: epidemiologic methods: statistics as topic: sensitivity and specificity research abstracts see: investigative techniques: epidemiologic methods: statistics as topic: sensitivity and specificity research

PUBMED ID PMID:

MEDLINE DATE:

A novel assay for monitoring internalization of nanocarrier coupled antibodies. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: BMC immunology

VOLUME: 7

Page Numbers: 24

Journal Abbreviation: BMC Immunol.

ISSN: 1471-2172

DAY: 2

MONTH: 10

YEAR: 2006

A novel assay for monitoring internalization of nanocarrier coupled antibodies. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 100966980

A novel assay for monitoring internalization of nanocarrier coupled antibodies. Keywords Mesh Terms:

KEYWORDS: Sensitivity and Specificity

MESH TERMS: immunology

Chemical & Substance for Abstract: A novel assay for monitoring internalization of nanocarrier coupled antibodies. Information

Substance Name: Receptor, erbB-2

Registry Number: EC 2.7.1.112

Grant and Affiliation Information for A novel assay for monitoring internalization of nanocarrier coupled antibodies.

AFFILIATION: Department of Anesthesia, University of California San Francisco, San Francisco, CA 94110, USA. unielsen@merrimackpharma.com

Country: England

AGENCY: United States NCI

GRANT: U54 CA90788

ACRONYM: CA

MEDLINETA: BMC Immunol

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

A novel assay for monitoring internalization of nanocarrier coupled antibodies Related Publications

• 5alpha-reductase: history and clinical importance.
• A novel assay for monitoring internalization of nanocarrier coupled antibodies.
• Anxiety sensitivity and auditory perception of heartbeat.
• Cannabis laws: an analysis of costs.
• Continuous participation in voluntary groups as a protective factor for the psychological well-being of adults who develop functional limitations: evidence from the national survey of families and households.
• Darkness descends with two Rabs.
• Evaluation of expression based markers for the detection of breast cancer cells.
• Genetic diversity among Botulinum Neurotoxin-producing clostridial strains.
• Granulocyte transfusions in the G-CSF era. Where do we stand?
• Histone deacetylase inhibitors selectively suppress expression of HDAC7.
• Histone deacetylase inhibitors: molecular mechanisms of action.
• Impact of single-chain Fv antibody fragment affinity on nanoparticle targeting of epidermal growth factor receptor-expressing tumor cells.
• Interrogational neuroimaging in counterterrorism: a "no-brainer" or a human rights hazard?
• MOSQUITOES (CULICIDAE) BITING A FISH (PERIOPHTHALMIDAE).
• Mechanisms of disease: Cytokine and adipokine signaling in uremic cachexia.
• Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin.
• Morbidity and cost of atopic eczema in Australia.
• One-pot terminal alkene homologation using a tandem olefin cross-metathesis/allylic carbonate reduction sequence.
• Penetrating spinal injury with wooden fragments causing cauda equina syndrome: case report and literature review.
• Predicting lung radiotherapy-induced pneumonitis using a model combining parametric Lyman probit with nonparametric decision trees.
• Recombinant full-length human IgG1s targeting hormone-refractory prostate cancer.
• Regulation of central melanocortin signaling by interleukin-1 beta.
• Self-consistent tumor control probability and normal tissue complication probability models based on generalized EUD.
• Skin cancer: a practical guide to management.
• The impact of induction chemotherapy and the associated tumor response on subsequent radiation-related changes in lung function and tumor response.
• The role of functional imaging in the diagnosis and management of late normal tissue injury.
• The subtypes of nicotinic acetylcholine receptors on dopaminergic terminals of mouse striatum.
• The way of all flesh - the 21st Century.
• Updated assessment of the six-minute walk test as predictor of acute radiation-induced pneumonitis.
• Who really wants to save the apes?