Application of Antibodies

Antibodies for Basic Research

Antibodies are indispensable molecules as tools for biological basic research. Many biotechnologies detect proteins or other molecules specifically by antibodies. Lots of common methods involve in antibodies such as flow cytometry, immunoblot, immunoprecipitation, enzyme-linked immunosorbent assay (ELISA), immunofluorescence, immunoprecipitation, purification of molecules or cells by affinity chromatography, phage display. In addition, research on the interaction between protein and protein or nucleic acid can achieve by the characteristics of antibody and antigen-specific binding, such as chromatin immunoprecipitation assay (ChIP) and co-Immunoprecipitation (CoIP). Many studies on protein molecular mechanisms are associated with modifications for proteins, such as phosphorylation, methylation, and acetylation, and these all require specific antibodies. Moreover, a significant application for antibodies in basic research is proteome research.

Antibodies for Diagnostics

Compared with other methods, antibody-based assays may provide a spatial, temporal, accurate, and quantitative measurement for the diagnosis of diseases. By the principle of specific binding of antigen-antibody, antibodies for diagnostics have already developed many products, especially in vitro diagnostic products. Now antibodies have a wide range of applications in medical diagnosis, which are used to detect pathogenic microbial and viral antigen, Parasite infection, antibody, tumor antigen, immunocytes and its subsets, hormones, and cytokines. There are three general ways to use antibodies for diagnostics: the detection of antigens using antibodies, the detection of serum antibodies using antigens, competition assay. The approaches of antibody-based diagnostic include ELISA, Mass Spectrometer (MS), real-time PCR, and diagnostic imaging techniques. But sample treatment is still needed in many cases, including expensive and complex laboratory devices.

Antibodies for Therapy

With the advent of advanced hybridoma techniques and phage display, antibodies are being investigated for several different applications in research and the clinic. Antibodies in the clinic have been used in versatile applications including, but not limited to, antibody-drug conjugates (ADCs), diagnostic agents, targeting ligands, and as therapeutics themselves. The antibody format has proven to be extremely amenable to protein engineering, which allows for modular design of structural domains that best integrate the desired therapeutic functions. For example, Small antigen-binding domains can be used to eliminate effector function, multimerizing antibodies increase complement fixation, and antibody fusions instill other proteins with favorable properties of the antibody framework. Conjugation of cytotoxic agents to antibodies allows for specific delivery of payloads to tumors, while multispecific antibodies grant novel mechanisms that increase specificity. Antibody engineering allows for the incorporation of amino acid and glycan changes that selectively alter biological and physical properties.

The first therapeutic monoclonal antibody, a murine anti-CD3 monoclonal antibody, was approved by the Food and Drug Administration (FDA) in 1985 to prevent the rejection of kidney, heart, and liver transplants. Generations of therapeutic antibodies have humanized the amino acid sequence of mouse antibodies to chimeric, humanized, and fully human. Mechanisms of therapeutic antibodies are manifold and include neutralization of substances, blocking of receptors, binding to cells and modulating the host immune system, or combinations of these effects. The most fruitful applications of antibodies lie in the fields of oncology (where built-in effector functions help to eliminate tumor cells) and immunology (where inhibition of inflammatory pathways is useful in treating autoimmunity). Antibodies used for cancer therapy involve direct antigen-mediated activity, checkpoint inhibition, targeting and recruitment of immune components such as NK cells, macrophages, or the complement system to cause cytotoxicity. In autoimmune diseases, the most widely used are anti-cytokine antibodies, in particular anti-TNFα, but also anti-CD20 and anti-CD25.

Therapeutic frameworks of antibodies. Fig1 Therapeutic frameworks of antibodies. (Goulet, 2019)

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  1. Goulet D.R.; et al. Considerations for the Design of Antibody-Based Therapeutics. Journal of Pharmaceutical Sciences. 2019, 109(1).

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