13 results on '"Wu, Anna M."'
Search Results
2. 18F-labeled anti-human CD20 cys-diabody for same-day immunoPET in a model of aggressive B cell lymphoma in human CD20 transgenic mice
- Author
-
Zettlitz, Kirstin A, Tavaré, Richard, Tsai, Wen-Ting K, Yamada, Reiko E, Ha, Noel S, Collins, Jeffrey, van Dam, R Michael, Timmerman, John M, and Wu, Anna M
- Subjects
Biomedical and Clinical Sciences ,Clinical Sciences ,Oncology and Carcinogenesis ,Rare Diseases ,Lymphoma ,Bioengineering ,Biomedical Imaging ,Cancer ,Hematology ,Orphan Drug ,4.1 Discovery and preclinical testing of markers and technologies ,Detection ,screening and diagnosis ,Animals ,Antibodies ,Antigens ,CD20 ,Fluorine Radioisotopes ,Humans ,Isotope Labeling ,Lymphoma ,B-Cell ,Mice ,Mice ,Transgenic ,Positron-Emission Tomography ,Radiochemistry ,Time Factors ,Tissue Distribution ,Anti-CD20 cys-diabody ,Antibody fragments ,ImmunoPET ,F-18 ,B cell lymphoma ,Same-day imaging ,18F ,Other Physical Sciences ,Nuclear Medicine & Medical Imaging ,Clinical sciences - Abstract
PurposeMetabolic imaging using [18F]FDG is the current standard for clinical PET; however, some malignancies (e.g., indolent lymphomas) show low avidity for FDG. The majority of B cell lymphomas express CD20, making it a valuable target both for antibody-based therapy and imaging. We previously developed PET tracers based on the humanised anti-CD20 antibody obinutuzumab (GA101). Preclinical studies showed that the smallest bivalent fragment, the cys-diabody (GAcDb, 54.5 kDa) with a peak uptake at 1-2 h post-injection and a biological half-life of 2-5 h, is compatible with short-lived positron emitters such as fluorine-18 (18F, t1/2 110 min), enabling same-day imaging.MethodsGAcDb was radiolabeled using amine-reactive N-succinimidyl 4-[18F]-fluorobenzoate ([18F]SFB), or thiol-reactive N-[2-(4-[18F]-fluorobenzamido)ethyl]maleimide ([18F]FBEM) for site-specific conjugation to C-terminal cysteine residues. Both tracers were used for immunoPET imaging of the B cell compartment in human CD20 transgenic mice (hCD20TM). [18F]FB-GAcDb immunoPET was further evaluated in a disseminated lymphoma (A20-hCD20) syngeneic for hCD20TM and compared to [18F]FDG PET. Tracer uptake was confirmed by ex vivo biodistribution.ResultsThe GAcDb was successfully 18F-radiolabeled using two different conjugation methods resulting in similar specific activities and without impairing immunoreactivity. Both tracers ([18F]FB-GAcDb and [18F]FBEM-GAcDb) specifically target human CD20-expressing B cells in transgenic mice. Fast blood clearance results in high contrast PET images as early as 1 h post injection enabling same-day imaging. [18F]FB-GAcDb immunoPET detects disseminated lymphoma disease in the context of normal tissue expression of hCD20, with comparable sensitivity as [18F]FDG PET but with added specificity for the therapeutic target.Conclusions[18F]FB-GAcDb and [18F]FBEM-GAcDb could monitor normal B cells and B cell malignancies non-invasively and quantitatively in vivo. In contrast to [18F]FDG PET, immunoPET provides not only information about the extent of disease but also about presence and localisation of the therapeutic target.
- Published
- 2019
3. Evaluation of [131I]I- and [177Lu]Lu-DTPA-A11 Minibody for Radioimmunotherapy in a Preclinical Model of PSCA-Expressing Prostate Cancer
- Author
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Tsai, Wen-Ting K, Zettlitz, Kirstin A, Dahlbom, Magnus, Reiter, Robert E, and Wu, Anna M
- Published
- 2020
- Full Text
- View/download PDF
4. Enhanced immunoPET of ALCAM-positive colorectal carcinoma using site-specific 64Cu-DOTA conjugation
- Author
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Tavaré, Richard, Wu, Wei H, Zettlitz, Kirstin A, Salazar, Felix B, McCabe, Katelyn E, Marks, James D, and Wu, Anna M
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Industrial Biotechnology ,Digestive Diseases ,Colo-Rectal Cancer ,Cancer ,Activated-Leukocyte Cell Adhesion Molecule ,Animals ,Cell Line ,Tumor ,Colorectal Neoplasms ,Copper ,Heterocyclic Compounds ,1-Ring ,Humans ,Immunohistochemistry ,Molecular Imaging ,Positron-Emission Tomography ,Rats ,Tissue Distribution ,ALCAM ,Cu-64 ,DOTA ,immunoPET ,site-specific conjugation ,Chemical Sciences ,Technology ,Biophysics ,Biochemistry and cell biology ,Industrial biotechnology - Abstract
Activated leukocyte cell adhesion molecule (ALCAM) is an immunoglobulin superfamily cell adhesion molecule that is aberrantly expressed in a wide variety of human tumors, including melanoma, prostate cancer, breast cancer, colorectal carcinoma, bladder cancer and pancreatic adenocarcinoma. This wide spectrum of human malignancies makes ALCAM a prospective pan-cancer immunoPET target to aid in detection and diagnosis in multiple malignancies. In this study, we assess site-specific versus non-site-specific conjugation strategies for (64)Cu-DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) immunoPET imaging of a fully human ALCAM cys-diabody (cDb) with a reduced linker length that retains its bivalent binding ability. ALCAM constructs with linker lengths of eight, five and three amino acids were produced to make true non-covalent site-specifically modified cDbs. Characterization by gel electrophoresis, size exclusion chromatography, flow cytometry and mass spectrometry of the various constructs was performed. To demonstrate the increased utility of targeting multiple malignancies expressing ALCAM, we compare the targeting of the site-specific versus non-site-specific conjugated cDbs to the human colorectal cancer xenograft LS174T. Interestingly, the conjugation strategy not only affects tumor targeting but also hepatic and renal uptake/clearance.
- Published
- 2014
5. ImmunoPET Imaging of Murine CD4+ T Cells Using Anti-CD4 Cys-Diabody: Effects of Protein Dose on T Cell Function and Imaging
- Author
-
Freise, Amanda C., Zettlitz, Kirstin A., Salazar, Felix B., Lu, Xiang, Tavaré, Richard, and Wu, Anna M
- Published
- 2017
- Full Text
- View/download PDF
6. F-18-labeled anti-human CD20 cys-diabody for same-day immunoPET in a model of aggressive B cell lymphoma in human CD20 transgenic mice
- Author
-
Zettlitz, Kirstin A, Tavare, Richard, Tsai, Wen-Ting K, Yamada, Reiko E, Ha, Noel S, Collins, Jeffrey, van Dam, R Michael, Timmerman, John M, and Wu, Anna M
- Subjects
Other Physical Sciences ,Nuclear Medicine & Medical Imaging ,18F ,Antibody fragments ,F-18 ,Clinical Sciences ,Same-day imaging ,Anti-CD20 cys-diabody ,ImmunoPET ,B cell lymphoma - Abstract
PURPOSE:Metabolic imaging using [18F]FDG is the current standard for clinical PET; however, some malignancies (e.g., indolent lymphomas) show low avidity for FDG. The majority of B cell lymphomas express CD20, making it a valuable target both for antibody-based therapy and imaging. We previously developed PET tracers based on the humanised anti-CD20 antibody obinutuzumab (GA101). Preclinical studies showed that the smallest bivalent fragment, the cys-diabody (GAcDb, 54.5 kDa) with a peak uptake at 1-2 h post-injection and a biological half-life of 2-5 h, is compatible with short-lived positron emitters such as fluorine-18 (18F, t1/2 110 min), enabling same-day imaging. METHODS:GAcDb was radiolabeled using amine-reactive N-succinimidyl 4-[18F]-fluorobenzoate ([18F]SFB), or thiol-reactive N-[2-(4-[18F]-fluorobenzamido)ethyl]maleimide ([18F]FBEM) for site-specific conjugation to C-terminal cysteine residues. Both tracers were used for immunoPET imaging of the B cell compartment in human CD20 transgenic mice (hCD20TM). [18F]FB-GAcDb immunoPET was further evaluated in a disseminated lymphoma (A20-hCD20) syngeneic for hCD20TM and compared to [18F]FDG PET. Tracer uptake was confirmed by ex vivo biodistribution. RESULTS:The GAcDb was successfully 18F-radiolabeled using two different conjugation methods resulting in similar specific activities and without impairing immunoreactivity. Both tracers ([18F]FB-GAcDb and [18F]FBEM-GAcDb) specifically target human CD20-expressing B cells in transgenic mice. Fast blood clearance results in high contrast PET images as early as 1 h post injection enabling same-day imaging. [18F]FB-GAcDb immunoPET detects disseminated lymphoma disease in the context of normal tissue expression of hCD20, with comparable sensitivity as [18F]FDG PET but with added specificity for the therapeutic target. CONCLUSIONS:[18F]FB-GAcDb and [18F]FBEM-GAcDb could monitor normal B cells and B cell malignancies non-invasively and quantitatively in vivo. In contrast to [18F]FDG PET, immunoPET provides not only information about the extent of disease but also about presence and localisation of the therapeutic target.
- Published
- 2019
7. Evaluation of [131I]I- and [177Lu]Lu-DTPA-A11 Minibody for Radioimmunotherapy in a Preclinical Model of PSCA-Expressing Prostate Cancer.
- Author
-
Tsai, Wen-Ting K, Zettlitz, Kirstin A, Dahlbom, Magnus, Reiter, Robert E, and Wu, Anna M
- Subjects
RADIOIMMUNOTHERAPY ,ANIMAL models in research ,PROSTATE cancer ,TUMOR growth ,STEM cells ,CYTOTOXIC T cells - Abstract
Purpose: Radioimmunotherapy uses tumor-specific antibodies to deliver therapeutic radionuclides, but hematological toxicity due to the long serum half-life of intact antibodies remains a challenge. We evaluated a smaller antibody fragment, the minibody, with faster kinetics and a potentially improved therapeutic index. Procedures: The anti-prostate stem cell antigen (PSCA) minibody (A11 Mb) was radiolabeled with iodine-124 ([
124 I]I-A11 Mb) or conjugated with deferoxamine (DFO) and labeled with zirconium-89 ([89 Zr]Zr-DFO-A11 Mb) for surrogate immunoPET to profile pharmacokinetics in a human prostate cancer xenograft model. Subsequently, minibodies labeled with two therapeutic beta emitters, directly iodinated [131 I]I-A11 Mb (non-residualizing) and177 Lu chelated using DTPA ([177 Lu]Lu-DTPA-A11 Mb) (residualizing), were compared for in vitro antigen-specific cytotoxicity. Full biodistribution studies (in 22Rv1-PSCA tumor bearing and hPSCA knock-in mice) were conducted for dosimetry calculations. Finally, the lead candidate [131 I]I-A11 Mb was evaluated in a radioimmunotherapy experiment. Escalating single doses (3.7, 11, or 37 MBq) and saline control were administered to 22Rv1-PSCA tumor bearing mice and anti-tumor effects (tumor volume) and toxicity (body weight) were monitored. Results: Minibodies radiolabeled with therapeutic beta emitters [131 I]I-A11 Mb and [177 Lu]Lu-DTPA-A11 Mb exhibited comparable tumor cell growth inhibition in vitro. In vivo surrogate immunoPET imaging using [89 Zr]Zr-DFO-A11 Mb showed activity retention in liver and kidney up to 72 h, while [124 I]I-A11 Mb cleared from liver, kidney, and blood by 48 h. Based on full biodistribution and dosimetry calculations, administering 37 MBq [131 I]I-A11 Mb was predicted to deliver a favorable dose to the tumor (35 Gy), with a therapeutic index of 22 (tumor:bone marrow). For [177 Lu]Lu-DTPA-A11 Mb, the kidneys would be dose-limiting, and the maximum tolerated activity (7.4 MBq) was not predicted to deliver an effective radiation dose to tumor. Radioimmunotherapy with a single dose of [131 I]I-A11 Mb showed dose-dependent tumor inhibition with minimal off-target toxicity and improved median survival (19 and 24 days, P < 0.001) compared with untreated mice (12 days). Conclusions: These findings show the potential of the anti-PSCA minibody for targeted radioimmunotherapy with minimal toxicity, and the application of immunoPET and dosimetry for personalized treatment. [ABSTRACT FROM AUTHOR]- Published
- 2020
- Full Text
- View/download PDF
8. Enhanced immunoPET of ALCAM-positive colorectal carcinoma using site-specific ⁶⁴Cu-DOTA conjugation
- Author
-
Tavaré, Richard, Wu, Wei H, Zettlitz, Kirstin A, Salazar, Felix B, McCabe, Katelyn E, Marks, James D, and Wu, Anna M
- Subjects
Technology ,Biophysics ,Cell Line ,Heterocyclic Compounds ,Activated-Leukocyte Cell Adhesion Molecule ,Animals ,Humans ,Tissue Distribution ,ALCAM ,Cancer ,1-Ring ,Tumor ,Cu-64 ,site-specific conjugation ,Biological Sciences ,Immunohistochemistry ,Rats ,Molecular Imaging ,Colo-Rectal Cancer ,DOTA ,Positron-Emission Tomography ,Chemical Sciences ,Colorectal Neoplasms ,immunoPET ,Digestive Diseases ,Copper - Abstract
Activated leukocyte cell adhesion molecule (ALCAM) is an immunoglobulin superfamily cell adhesion molecule that is aberrantly expressed in a wide variety of human tumors, including melanoma, prostate cancer, breast cancer, colorectal carcinoma, bladder cancer and pancreatic adenocarcinoma. This wide spectrum of human malignancies makes ALCAM a prospective pan-cancer immunoPET target to aid in detection and diagnosis in multiple malignancies. In this study, we assess site-specific versus non-site-specific conjugation strategies for (64)Cu-DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) immunoPET imaging of a fully human ALCAM cys-diabody (cDb) with a reduced linker length that retains its bivalent binding ability. ALCAM constructs with linker lengths of eight, five and three amino acids were produced to make true non-covalent site-specifically modified cDbs. Characterization by gel electrophoresis, size exclusion chromatography, flow cytometry and mass spectrometry of the various constructs was performed. To demonstrate the increased utility of targeting multiple malignancies expressing ALCAM, we compare the targeting of the site-specific versus non-site-specific conjugated cDbs to the human colorectal cancer xenograft LS174T. Interestingly, the conjugation strategy not only affects tumor targeting but also hepatic and renal uptake/clearance.
- Published
- 2014
9. 18F-labeled anti-human CD20 cys-diabody for same-day immunoPET in a model of aggressive B cell lymphoma in human CD20 transgenic mice.
- Author
-
Tsai, Wen-Ting K., Collins, Jeffrey, Zettlitz, Kirstin A., Wu, Anna M., Tavaré, Richard, Ha, Noel S., van Dam, R. Michael, Yamada, Reiko E., and Timmerman, John M.
- Subjects
LYMPHOMAS ,CD20 antigen ,IMMUNOGLOBULINS ,POSITRON emission ,FLUORODEOXYGLUCOSE F18 ,GENE expression ,DIAGNOSTIC imaging - Abstract
Purpose: Metabolic imaging using [
18 F]FDG is the current standard for clinical PET; however, some malignancies (e.g., indolent lymphomas) show low avidity for FDG. The majority of B cell lymphomas express CD20, making it a valuable target both for antibody-based therapy and imaging. We previously developed PET tracers based on the humanised anti-CD20 antibody obinutuzumab (GA101). Preclinical studies showed that the smallest bivalent fragment, the cys-diabody (GAcDb, 54.5 kDa) with a peak uptake at 1-2 h post-injection and a biological half-life of 2-5 h, is compatible with short-lived positron emitters such as fluorine-18 (18 F, t1/2 110 min), enabling same-day imaging.Methods: GAcDb was radiolabeled using amine-reactive N-succinimidyl 4-[18 F]-fluorobenzoate ([18 F]SFB), or thiol-reactive N-[2-(4-[18 F]-fluorobenzamido)ethyl]maleimide ([18 F]FBEM) for site-specific conjugation to C-terminal cysteine residues. Both tracers were used for immunoPET imaging of the B cell compartment in human CD20 transgenic mice (hCD20TM). [18 F]FB-GAcDb immunoPET was further evaluated in a disseminated lymphoma (A20-hCD20) syngeneic for hCD20TM and compared to [18 F]FDG PET. Tracer uptake was confirmed by ex vivo biodistribution.Results: The GAcDb was successfully18 F-radiolabeled using two different conjugation methods resulting in similar specific activities and without impairing immunoreactivity. Both tracers ([18 F]FB-GAcDb and [18 F]FBEM-GAcDb) specifically target human CD20-expressing B cells in transgenic mice. Fast blood clearance results in high contrast PET images as early as 1 h post injection enabling same-day imaging. [18 F]FB-GAcDb immunoPET detects disseminated lymphoma disease in the context of normal tissue expression of hCD20, with comparable sensitivity as [18 F]FDG PET but with added specificity for the therapeutic target.Conclusions: [18 F]FB-GAcDb and [18 F]FBEM-GAcDb could monitor normal B cells and B cell malignancies non-invasively and quantitatively in vivo. In contrast to [18 F]FDG PET, immunoPET provides not only information about the extent of disease but also about presence and localisation of the therapeutic target. [ABSTRACT FROM AUTHOR]- Published
- 2019
- Full Text
- View/download PDF
10. Aligning physics and physiology: Engineering antibodies for radionuclide delivery.
- Author
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Tsai, Wen‐Ting K. and Wu, Anna M.
- Subjects
- *
CANCER treatment , *RADIOIMMUNOTHERAPY , *RADIOISOTOPES , *IMMUNOTECHNOLOGY , *RADIOLABELING - Abstract
The exquisite specificity of antibodies and antibody fragments renders them excellent agents for targeted delivery of radionuclides. Radiolabeled antibodies and fragments have been successfully used for molecular imaging and radioimmunotherapy (RIT) of cell surface targets in oncology and immunology. Protein engineering has been used for antibody humanization essential for clinical applications, as well as optimization of important characteristics including pharmacokinetics, biodistribution, and clearance. Although intact antibodies have high potential as imaging and therapeutic agents, challenges include long circulation time in blood, which leads to later imaging time points post‐injection and higher blood absorbed dose that may be disadvantageous for RIT. Using engineered fragments may address these challenges, as size reduction and removal of Fc function decreases serum half‐life. Radiolabeled fragments and pretargeting strategies can result in high contrast images within hours to days, and a reduction of RIT toxicity in normal tissues. Additionally, fragments can be engineered to direct hepatic or renal clearance, which may be chosen based on the application and disease setting. This review discusses aligning the physical properties of radionuclides (positron, gamma, beta, alpha, and Auger emitters) with antibodies and fragments and highlights recent advances of engineered antibodies and fragments in preclinical and clinical development for imaging and therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
11. ImmunoPET Imaging of Murine CD4+ T Cells Using Anti-CD4 Cys-Diabody: Effects of Protein Dose on T Cell Function and Imaging.
- Author
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Freise, Amanda, Zettlitz, Kirstin, Salazar, Felix, Lu, Xiang, Tavaré, Richard, Wu, Anna, Freise, Amanda C, Zettlitz, Kirstin A, Salazar, Felix B, Tavaré, Richard, and Wu, Anna M
- Subjects
POSITRON emission tomography ,CELL imaging ,T cells ,CD4 antigen ,MOLECULAR probes ,CANCER immunotherapy - Abstract
Purpose: Molecular imaging of CD4+ T cells throughout the body has implications for monitoring autoimmune disease and immunotherapy of cancer. Given the key role of these cells in regulating immunity, it is important to develop a biologically inert probe. GK1.5 cys-diabody (cDb), a previously developed anti-mouse CD4 antibody fragment, was tested at different doses to assess its effects on positron emission tomography (PET) imaging and CD4+ T cell viability, proliferation, CD4 expression, and function.Procedures: The effect of protein dose on image contrast (lymphoid tissue-to-muscle ratio) was assessed by administering different amounts of 89Zr-labeled GK1.5 cDb to mice followed by PET imaging and ex vivo biodistribution analysis. To assess impact of GK1.5 cDb on T cell biology, GK1.5 cDb was incubated with T cells in vitro or administered intravenously to C57BL/6 mice at multiple protein doses. CD4 expression and T cell proliferation were analyzed with flow cytometry and cytokines were assayed.Results: For immunoPET imaging, the lowest protein dose of 2 μg of 89Zr-labeled GK1.5 cDb resulted in significantly higher % injected dose/g in inguinal lymph nodes (ILN) and spleen compared to the 12-μg protein dose. In vivo administration of GK1.5 cDb at the high dose of 40 μg caused a transient decrease in CD4 expression in spleen, blood, lymph nodes, and thymus, which recovered within 3 days postinjection; this effect was reduced, although not abrogated, when 2 μg was administered. Proliferation was inhibited in vivo in ILN but not the spleen by injection of 40 μg GK1.5 cDb. Concentrations of GK1.5 cDb in excess of 25 nM significantly inhibited CD4+ T cell proliferation and interferon-γ production in vitro. Overall, using low-dose GK1.5 cDb minimized biological effects on CD4+ T cells.Conclusions: Low-dose GK1.5 cDb yields high-contrast immunoPET images with minimal effects on T cell biology in vitro and in vivo and may be a useful tool for investigating CD4+ T cells in the context of preclinical disease models. Future approaches to minimizing biological effects may include the creation of monovalent fragments or selecting anti-CD4 antibodies which target alternative epitopes. [ABSTRACT FROM AUTHOR]- Published
- 2017
- Full Text
- View/download PDF
12. In vivo imaging with antibodies and engineered fragments.
- Author
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Freise, Amanda C. and Wu, Anna M.
- Subjects
- *
IMMUNOGLOBULINS , *HEMATOLOGY , *DRUG efficacy , *COMMUNICABLE diseases , *IMAGING systems , *NONINVASIVE diagnostic tests - Abstract
Antibodies have clearly demonstrated their utility as therapeutics, providing highly selective and effective drugs to treat diseases in oncology, hematology, cardiology, immunology and autoimmunity, and infectious diseases. More recently, a pressing need for equally specific and targeted imaging agents for assessing disease in vivo , in preclinical models and patients, has emerged. This review summarizes strategies for developing and optimizing antibodies as targeted probes for use in non-invasive imaging using radioactive, optical, magnetic resonance, and ultrasound approaches. Recent advances in engineered antibody fragments and scaffolds, conjugation and labeling methods, and multimodality probes are highlighted. Importantly, antibody-based imaging probes are seeing new applications in detection and quantitation of cell surface biomarkers, imaging specific responses to targeted therapies, and monitoring immune responses in oncology and other diseases. Antibody-based imaging will provide essential tools to facilitate the transition to truly precision medicine. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
13. Engineered antibodies for molecular imaging of cancer.
- Author
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Wu, Anna M.
- Subjects
- *
IMAGING of cancer , *CANCER immunotherapy , *MONOCLONAL antibodies , *CANCER diagnosis , *PHARMACOKINETICS , *POSITRON emission tomography - Abstract
Abstract: Antibody technology has transformed drug development, providing robust approaches to producing highly targeted and active therapeutics that can routinely be advanced through clinical evaluation and registration. In parallel, there is an emerging need to access similarly targeted agents for diagnostic purposes, including non-invasive imaging in preclinical models and patients. Antibody engineering enables modification of key properties (immunogenicity, valency, biological inertness, pharmacokinetics, clearance route, site-specific conjugation) in order to produce targeting agents optimized for molecular imaging. Expanded availability of positron-emitting radionuclides has led to a resurgence of interest and applications of immunoPET (immuno-positron emission tomography). Molecular imaging using engineered antibodies and fragments provides a general approach for assessing cell surface phenotype in vivo and stands to play an increasingly important role in cancer diagnosis, treatment selection, and monitoring of molecularly targeted therapeutics. [Copyright &y& Elsevier]
- Published
- 2014
- Full Text
- View/download PDF
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