免疫学技术

Virology has greatly benefited from the introduction of fluorescent proteins (FP’s) as tags to viral as well as cellular structures. With advanced imaging technologies it is now possible to observe host–pathogen interactions in living cell systems in real-time. The generation of high- ...

Fluorescence in situ hybridization (FISH) is widely used to visualize nucleotide sequences in interphase cells or on metaphase chromosomes using specific probes that are complementary to the respective targets. Besides its broad application in cytogenetics and cancer researc ...

Multimeric antibody fragments, particularly dimers (diabodies), trimers (triabodies), and tetramers (tetrabodies) of single-chain Fv molecules (scFv), provide high avidity through multivalent binding to the target antigen. The combination of their smaller size and avid bin ...

Radiolabeled antibodies were studied first for tumor detection by single-photon imaging, but FDG PET stopped these developments. In the meantime, radiolabeled antibodies were shown to be effective in the treatment of lymphoma. Radiolabeling techniques are well established and r ...

Intracellularly expressed recombinant antibodies, or intrabodies, are powerful tools for cell biology studies as well as therapeutic applications. Cell biologists use them to either block the intracellular antibody target or to image endogenous target dynamics. We describe he ...

The Chimeric Antigen Receptor (CAR) consists of an antibody-derived targeting domain fused with T-cell signaling domains that, when expressed by a T-cell, endows the T-cell with antigen specificity determined by the targeting domain of the CAR. CARs can potentially redirect the effector ...

Recombinant immunotoxin (RIT) contains the Fv portion of the antibody fused to the truncated form of toxin and are ongoing in clinical trials for cancer therapy. To obtain high yields of products, RITs are produced in Escherichia coli (E. coli). As the endotoxin came from E. coli cells and is harmful to an ...

Increasing insight into the misbalance and poor activity of tumor infiltrating immune cells raised interest to activate and improve an antitumor immune response by accumulating IL2 in the tumor tissue. IL2 can be targeted as part of an antibody-cytokine fusion protein to the tumor tissue by a s ...

Single-chain variable fragments (scFvs) specific for tumor-associated cell surface antigens are the most broadly used reagents to direct therapeutic or diagnostic effector molecules, such as toxins, radioisotopes, and CD3-stimulating scFvs, to tumors. One novel class of effec ...

This chapter describes in vitro and in vivo methods to characterize a lead monoclonal antibody candidate in the drug discovery setting. Approaches to characterize monoclonal antibody specificity, heavy and light chain composition, and antibody mode of action including the ability ...

Controlling the half-life of pharmaceuticals through Fc engineering is a desirable approach to achieve optimal exposure and targeting. The long serum residence time of gamma immunoglobulins is attributed to the Fc binding to the neonatal Fc receptor (FcRn). The residues in the Fc region th ...

Today monoclonal antibodies are widely used in cancer therapy. However, clinical experience as well as translational research into antibodies’ pharmacology and effector mechanisms has identified limitations of antibody therapy, including inefficient effector cell recr ...

Glycosylation is known to have an impact on pharmacokinetics and pharmacodynamics of therapeutic proteins. While the production of pharmaceutically desirable glycosylation forms of a therapeutic protein can in certain cases be influenced by the upstream process parameters, c ...

Protein engineering techniques can facilitate the direct de-convolution of specific domains, regions, and particular amino acids that contribute to protein function. Many tools are available to aid this enterprise and herein we describe one such tool, a technique we term “Molecular S ...

Rational engineering methods can be applied with success to optimize physicochemical characteristics of antibodies. Application of in silico analysis and prediction methods to antibody Fv regions can help to find residues affecting antibody–antigen affinity when high-res ...

As a growing number of therapeutic antibodies are developed, robust methods to efficiently improve the affinity and/or specificity of antibody candidates are needed. Here we describe our powerful platform that combines scFv affinity maturation and IgG high-throughput screenin ...

Affinity determination is a crucial step of an antibody characterization. Here, we describe a method for antibody affinity determination by flow cytometry, relying on the unique affinity of biotin for streptavidin for easy and efficient antibody labeling. Several labeling approac ...

Surface plasmon resonance (SPR) is now widely embraced as a technology for monitoring a diverse range of protein–protein interactions and is considered almost de rigueur for characterizing antibody–antigen interactions. The technique obviates the need to label either of the inter ...

Expression and purification of recombinant proteins produced in plants is emerging as an affordable alternative to using more costly mammalian bioreactors since plants are capable of producing mammalian proteins at high concentrations. There are two general methods of express ...

The baculovirus–insect cell system is effective in the production of large quantities of biologically active recombinant proteins. The secretory production of an antibody Fab fragment in lepidopteran insect cells infected with a recombinant baculovirus that contains both the h ...