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Protocol
Immunophenoty**, Part II: Analysis of Nanoparticle Effects on the Composition and Activation Status of Human Peripheral Blood Mononuclear Cells
The use of nanoparticles as drug delivery carriers requires analysis of their safety, which among other tests, includes immunotoxicity. Nanoparticles are also increasingly used for applications intended to spe...
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Protocol
Immunophenoty**, Part I: Instrument Calibration and Reagent Qualification for Immunophenoty** Analysis of Human Peripheral Blood Mononuclear Cell Cultures
Nanoparticles are increasingly used in biomedical applications to influence the way the immune system reacts to tumors and infectious disease-causing agents. Nanoparticles not-intended for immunomodulation can...
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Protocol
Characterizing the In Vivo Role of Candidate Leukemia Stem Cell Genes
Acute myeloid leukemia (AML) is a disease caused by multiple distinct genomic events in the hematopoietic stem cell and progenitor compartment. To gain insight into the link between genetic mutations in AML an...
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Protocol
Characteristics of Major Drug Metabolizing Cytochrome P450 Enzymes
Cytochrome P450 (CYP) enzymes have been widely studied for drug discovery and development. While much has been discovered in terms of characteristics of each major drug metabolizing CYPs, challenges still exis...
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Protocol
Self-Assembling Peptide Nanofibrous Scaffolds in Central Nervous System Lesions
Central nervous system (CNS) lesion leads to the loss of tissues and functional defect. After injury, the local microenvironment typically contains potential inhibitory molecules and glial scars, which inhibit...
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Protocol
Deep Learning-Based Drug Screening for COVID-19 and Case Studies
Coronavirus infectious disease (COVID-19), caused by deadly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been declared as a pandemic by the World Health Organization. This disease has beco...
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Protocol
Conjugation Site Analysis by MS/MS Protein Sequencing
In-depth knowledge about the site of drug-linker conjugation is important for the understanding of the conjugation efficiency and the exact locations of payloads for antibody-drug conjugates (ADCs). Here we de...
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Protocol
Fast and Efficient Measurement of Clinical and Biological Samples Using Immunoassay-Based Multiplexing Systems
Immunoassay is one of the most commonly used biomedical techniques to detect the expression of an antibody or an antigen in a test sample. Enzyme-linked immunosorbent assay (ELISA) has been used for a variety ...
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Protocol
Analysis of Mutations in K-ras and p53 Genes in Sputum and Plasma Samples
Sputum and plasma can provide noninvasive materials to investigate biomarkers for cancer detection and diagnosis. Mutations in the K-ras oncogene and p53 suppressor gene have been frequently found in sputum an...
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Protocol
Testing of Schistosoma mansoni Vaccine Targets
Vaccination has greatly reduced the burden of human diseases caused by infectious pathogens. Systematic development of vaccine targets requires established protocols to assess immunogenicity and efficacy of su...
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Protocol
Application of Gelatin Zymography in Nanotoxicity Research
Gelatin zymography is a relatively simple, inexpensive, and powerful technique to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It has been used particularly to detec...
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Protocol
Evaluation of Pulmonary Toxicity of Nanoparticles by Bronchoalveolar Lavage
Nanoparticles can enter the body via the lungs, the digestive tract, and the skin. The pulmonary effects of nanoparticles and the underlying mechanisms may be investigated by histopathological examination, imm...
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Protocol
SpyTag–SpyCatcher Chemistry for Protein Bioconjugation In Vitro and Protein Topology Engineering In Vivo
The emergence of “molecular superglue,” such as SpyTag–SpyCatcher chemistry, has tremendously expanded our capability in manipulating protein shape and architecture via conjugation. Telechelic proteins bearing...
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Protocol
Site-Selective Enzymatic Labeling of Designed Ankyrin Repeat Proteins Using Protein Farnesyltransferase
Affinity agents coupled to a functional moiety play an ever-increasing role in modern medicine, ranging from radiolabeled selective binders in diagnosis to antibody–drug conjugates in targeted therapies. In bi...
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Protocol
Analysis of Nanomaterial Toxicity by Western Blot
Western blot is a routine biochemical technique for the immunodetection of proteins in cells and tissues exposed to nanomaterials (NMs). It is a sensitive method for protein analysis that involves the solubili...
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Protocol
Methods for Evaluation of Ovarian Granulosa Cells with Exposure to Nanoparticles
Increasing evidence demonstrates that a substantial number of chemicals including nanoparticles have potential toxic effects on human and wildlife reproductive health. Ovarian granulosa cells (GCs), the functi...
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Protocol
Determination of Phosphorylated Histone H2AX in Nanoparticle-Induced Genotoxic Studies
DNA double-strand breaks (DSBs), one of the most severe lesions of DNA damage triggered by various genotoxic insults, can lead to chromosome change, genomic instability, and even tumorigenesis if not repaired ...
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Protocol
Application of Electron Microscopes in Nanotoxicity Assessment
In this chapter, we highlight the applications of electron microscopes (EMs) in nanotoxicity assessment. EMs can provide detailed information about the size and morphology of nanomaterials (NMs), their localiz...
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Protocol
The Overview of Methods of Nanoparticle Exposure Assessment
Nanotechnology is now widely used in industry as well as consumer products, such as electrical devices, cosmetics, medicine, and household appliances. In the life cycle of the nano-products, including producti...
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Protocol
NIR Imaging-Guided Photothermal Therapy by cRGD-Conjugated Solid Lipid Nanoparticles Encapsulating IR-780
It is extremely demanded to enhance the accumulation of near-infrared theranostic agents in the tumor region, which is favorable to the effective phototherapy. Compared with indocyanine green (a clinically app...