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Han et al. first described in vitro bioconjugation of a full-length bispecific antibody via intein fusions to precursor antibody fragments within the hinge region and successfully demonstrated in vivo activity of a reconstituted CD3xHer2 T-cell engager mediated by Npu DnaE . A similar approach yielded a CD3xPRLR bispecific antibody for T-cell activation and cytokine release towards PRLR expressing breast cancer cells . In addition to Fc-based bispecifics, also non-Fc, circularly connected VHH fragments (cyclobody) have been developed via SICLOPPS (Split Intein Circular Ligation of Peptides and Proteins) reaction between both C- and N-termini, forming a cyclic conformation after PTS . These cyclobodies are, as discussed before for other applications, protected from proteolysis due to their cyclic topology, yet they retain their dual specificity. An anti-EGFRxCD16 cyclobody was successfully generated to show cytotoxicity against EGFR-positive cancer cells, able to bind simultaneously EGFR and CD16 on the cell surface . Applicability of the aforementioned split intein Aes PolB1 intein was demonstrated by successful bioconjugation of several therapeutically relevant formats like full-length IgG, Fc, and VHH fusions . Split inteins have been used to bioconjugate toxic components to antibodies , avoiding toxicity issues during antibody production, as exemplified by an anti-Her2 immunotoxin conjugated via split intein derivative (M86) of the Ssp DnaB intein . Next to several split inteins applied in bioconjugation of single binders in diverse formats, we recently described the application of split inteins for automated high throughput screenings in pharmaceutical research and development . Similar to other presented methodologies, split intein screening could enable the comparison of complex formats with feasible low production needs and faster development times.
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Bioconjugate Chemistry invites original contributions on all aspects of the joining of different molecular functions by chemical or biological means. This includes, among other topics, the conjugation of antibodies, nucleic acids, lipids, carbohydrates, or other biologically active molecules and their analogs with any molecular groups that add useful properties (drugs, radionuclides, toxins, fluorophores, photoprobes, inhibitors, enzymes, haptens, ligands, etc.). Topics of current interest include molecules that influence cellular uptake and trafficking and also those attached to particles or surfaces. Bioconjugate Chemistry is intended to provide a forum for presentation of research relevant to all aspects of conjugation chemistry and biochemistry, including the preparation, characterization, and properties (both chemical and biological) of molecular conjugates. The journal emphasizes rigorous chemical standards and encourages application of modern techniques of chemical analysis to problems in conjugation chemistry. Read Less
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In the years since the publication of the second edition, the field of bioconjugation has continued to advance at an incredible pace. Since 2008, over 54,000 additional journal publications have appeared in the biological, medical, polymer, material science, and chemistry journals that at least mention the terms bioconjugate or bioconjugation. In addition, many tens of thousands of new links to Internet sites with bioconjugation information also have appeared in this time frame, including sources from academic, corporate, and personal web pages. These journal articles and links describe many new reagents and reactions for forming bioconjugates of all types, including the formation of unique complexes in solution as well as the coupling of molecules to solid-phase surfaces or particles. In addition, exciting new methods are appearing for the application of bioconjugates in highly sensitive assays and detection schemes, for in vivo imaging and diagnosis, for therapeutic drug targeting, in the capture and purification of biomolecules, for catalysis and chemical modification, and for vaccine development and immune modulation. These recent advances in bioconjugate techniques have resulted in two new chapters and many new sections and updates throughout the book, as well as the rearrangement and consolidation of chapters to more logically group topics together having common themes.
The third edition also contains two major chapters that were obvious gaps in the previous editions: Chapter 1 is an extensive introduction to the vast field of bioconjugation, while Chapter 15 describes the reagents and techniques used for the immobilization of ligands onto chromatography supports. The new comprehensive introduction to the book begins by describing the basic principles of bioconjugation along with presenting important strategies for designing optimal conjugates for a wide range of applications. Chapter 1 also reviews the major application areas where bioconjugates are being used today and describes the conjugate designs associated with each of these applications.
Another major change that is immediately noticeable with this edition is the use of full-color illustrations. Literally hundreds of new and updated figures now use color to better illustrate reactions or to show how bioconjugates are being used in applications. While the design of the book may have been radically changed and updated with this edition, it is my hope that the reader will continue to find it useful in the design of new bioconjugates.
This book describes hundreds of reagents, reactions, and applications for use in bioconjugation. Most of the compounds are highly specialized and we have very little information regarding their toxicological properties. At a minimum, bioconjugation reagents should be considered irritants and handled with care. However, the overwhelming majority are known to be reactive and any individual compound or solvent can be corrosive, hazardous, toxic, volatile, flammable, explosive, or otherwise dangerous to personal health and safety. For this reason, the use of any reagent or protocol described in this book should be carried out by taking the appropriate precautions. Before utilizing any of these methods, the user agrees to take complete responsibility and personal liability for any and all risks associated with the reagents and reactions described in this book or within the references cited. Before starting an experiment, the user agrees to reference the appropriate Material Safety Data Sheets (MSDS) relative to every compound or component used in a reaction and to completely understand the properties of the reactions being contemplated. The use of personal protective equipment (PPE), fume hoods, and proper laboratory techniques can ensure safety for both the user and other people in the immediate vicinity. In addition, the disposal of waste materials should be performed according to the appropriate environmental regulations to prevent toxic elements or compounds from entering the water, air, or soil. The inappropriate disposal of excess reagents or reaction byproducts may be harmful to people and the environment.
Throughout this book I have provided references related to the reagents, reactions, and techniques used in bioconjugation. There are many additional references that can be found by performing the appropriate key word searches on the Internet. However, such knowledge does not necessarily provide the liberty to legally use these reagents and applications for commercial purposes without consideration for existing intellectual property rights. While in some cases pertinent patent references are provided within the book, this is done only to supply additional technical details about the topic being discussed and not to imply anything about freedom to operate.
Today, nearly every important reagent or method reported in the literature has a patent or patent application associated with it, especially if it has potential commercial value. A search of the patent databases, such as the United States Patent and Trademark Office ( ) or the European Patent Office ( ) for key words or the names of inventors can provide a list of existing issued patents or patent applications related to a bioconjugate technique or compound. 350c69d7ab