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丁香实验推荐阅读
Risk-Based Scientific Approach for Determination of Extractables/Leachables from Biomanufacturing of AntibodyDrug Conjugates (ADCs)

Recent developments in biopharmaceutical processes twined with a desire to remove cleaning and cross-contamination issues from drug production have led to the widespread introduction of single-use technologies and systems within operations. One key area that end users need to ad ...

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AntibodyDrug Conjugate (ADC) Clinical Pipeline: A Review

Biological therapies play an increasing role in cancer treatment, although the number of naked antibodies showing clinical efficacy as single agent remains limited. One way to enhance therapeutic potential of antibodies is to conjugate them to small molecule drugs. This combination ...

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Determination of Charge Heterogeneity and Level of Unconjugated Antibody by Imaged cIEF

Imaged capillary isoelectric focusing (icIEF) is capable of monitoring the charge heterogeneity profile of conjugated antibodies. The electropherogram from icIEF can be integrated to quantitate the amount of unconjugated antibody present in a conjugate sample. This chapter d ...

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Drug-to-Antibody Ratio (DAR) and Drug Load Distribution by LC-ESI-MS

This chapter describes an LC-ESI-MS method for the DAR and drug load distribution analysis that is suitable for lysine-linked ADCs. The ADC sample is desalted using a reversed-phase LC column with an acetonitrile gradient prior to online MS analysis. The MS spectrum is processed (deconvolut ...

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Drug-to-Antibody Ratio (DAR) and Drug Load Distribution by Hydrophobic Interaction Chromatography and Reversed Phase High-Performance Liquid Chromatog

Hydrophobic interaction chromatography (HIC) is the method of choice for determination of the drug-to-antibody ratio (DAR) and drug load distribution for cysteine (Cys)-linked antibody–drug conjugates (ADCs). The drug-loaded species are resolved based on the increasing hydro ...

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Drug-to-Antibody Ratio (DAR) by UV/Vis Spectroscopy

UV/Vis spectroscopy is a simple and convenient method to determine protein concentrations as well as the average number of drugs that are conjugated to the antibody in an antibody–drug conjugate (ADC). Using the measured absorbances of the ADC and the extinction coefficients of the antibody ...

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Methods for Conjugating Antibodies to Nanocarriers

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 effic ...

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Conjugation Process Development and Scale-Up

Manufacturing highly potent antibody–drug conjugates (ADCs) is a demanding task—combining conventional organic synthesis with biotechnological manufacturing. Hence a series of new and unique engineering and chemistry challenges have to be addressed to support clinical t ...

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Formulation Development of AntibodyDrug Conjugates

Formulation development of an ADC resembles that of a conventional antibody, but the conjugated form introduces new molecular attributes such as drug-to-antibody ratio and stability of the drug itself that need to be considered. An extended set of analytical tools, coupled with underst ...

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Enzymatic Antibody Modification by Bacterial Transglutaminase

Enzymatic posttranslational modification of proteins permits more precise control over conjugation site than chemical modification of reactive amino acid side chains. Ideally, protein modification by an enzyme yields completely homogeneous conjugates with improved pr ...

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Engineering THIOMABs for Site-Specific Conjugation of Thiol-Reactive Linkers

Antibody conjugates are used in many therapeutic and research applications and are generated by chemically linking a cysteine or lysine residue to potent chemotherapeutic drugs or other functional groups through a flexible linker. Recently, we have engineered THIOMABs (antibod ...

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Protocols for Lysine Conjugation

Currently, the most widely used chemical methodology for the conjugation of drugs to monoclonal antibodies involves either lysine or cysteine residues. In this chapter, several methods for the preparation of antibody–drug conjugates (ADCs) through conjugation of drugs to solvent ...

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Genotoxicity Guidelines Recommended by International Conference of Harmonization (ICH)

Genotoxicity tests are designed to detect the genetic damage by various mechanisms. Several guidelines have provided various tests to be conducted for testing the genotoxicity and each of the regulatory agencies around the world have developed their own requirements for mutagenic ...

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The Wing-Spot and the Comet Tests as Useful Assays Detecting Genotoxicity in Drosophila

In spite of its pioneer use in detecting mutational processes, Drosophila has yet an important role in studies aiming to detect and quantify the induction of DNA damage. Here we describe two assays, one detecting primary damage (the Comet assay) and the other detecting somatic mutation and recom ...

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Micronucleus Assay with Tetrad Cells of Tradescantia

The Tradescantia micronucleus assay is being used since almost 50 years for the detection of genotoxins (including carcinogens) in the environment. A large database on the effects of individual compounds and of complex environmental mixtures (soil, air and water) has accumulated. In co ...

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Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells In Vivo

The unscheduled DNA synthesis test is a short-term genotoxicity assay that allows identification of substances that induce DNA repair in liver cells of treated animals. This endpoint can be assessed by determining the uptake of radiolabelled nucleotide after excision-repair in cel ...

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The Comet Assay in Marine Animals

Comet assay is a quick and versatile technique for assessing DNA damage in individual cells. It allows the detection of DNA single- and double-strand breaks, as well as the presence of alkali-labile sites and cross-links. Here we describe the protocols for the single-cell gel electrophoresis ( ...

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The Comet Assay in Human Biomonitoring

Human biomonitoring studies aim to identify potential exposures to environmental, occupational, or lifestyle toxicants in human populations and are commonly used by public health decision makers to predict disease risk. The Comet assay measures changes in genomic stability and is ...

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The Comet Assay: Assessment of In Vitro and In Vivo DNA Damage

Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organi ...

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Multicolor Laser Scanning Confocal Immunofluorescence Microscopy of DNA Damage Response Biomarkers

DNA damage through endogenous and environmental toxicants is a constant threat to both a human’s ability to pass on intact genetic information to its offspring as well as somatic cells for their own survival. To counter these threats posed by DNA damage, cells have evolved a series of highly choreo ...

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