Bioconjugation Protocols

There are a number of outstanding volumes that provide a comprehensive overview of bioconjugation techniques. However, many of the conventional approaches to the synthesis of chemically modified protein conjugates lack efficient means to control the stoichiometry of conjugation, as well as the s- cific site of attachment of the conjugated moiety. Moreover, the recent dev- opments in microarray technologies as well as in nanobiotechnology—a novel field of research rapidly evolving at the crossroads of physics, chemistry, b- technology, and materials science—call for a summary of modern bioconjugation strategies to overcome the limitations of the classical approaches. Bioconjugation Protocols: Methods and Strategies is intended to provide an update of many of the classic techniques and also to introduce and summarize newer approaches that go beyond the pure biomedical applications of bioconjugation. The purpose of Bioconjugation Protocols: Methods and Str- egies is therefore to provide instruction and inspiration for all those scientists confronting the challenges of semisynthesizing functional biomolecular reagents for a wide variety of applications ranging from novel biomedical diagnostics, to therapeutics, to biomaterials. Part I contains seven protocols for the preparation of protein conjugates.

Contemporary approaches to the synthesis of chemically modified biomacromolecules (proteins, nucleic acids, lipids, and carbohydrates) not only require efficient means to control conjugation and the specific site of attachment of the conjugated moiety but also the effective use of recent developments in the fields of pharmaceutical chemistry, biomolecular/polymer engineering, and nanobiotechnology. In this second edition of Bioconjugation Protocols: Strategies and Methods, expert researchers update the classic methods and introduce valuable new approaches that go beyond basic conjugation techniques to include elements from advanced organic synthesis, molecular biology, surface biotechnology, materials science, and nanobioscience/engineering. These readily reproducible methods cover the preparation of biomolecular conjugates using a variety of labeling techniques and semisynthetic approaches. Additional chapters address the biofunctionalization of surface structures, including organic/inorganic thin films, as well as various types of nanostructures (magnetic nanoparticles, quantum dots, carbon nanotubes, and silicon nanowire devices). All the protocols follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and highly practical, Bioconjugation Protocols: Strategies and Methods, Second Edition offers both novice and experienced researchers access to the broad array of techniques needed to carry out the semisynthesis of functional biomolecular reagents and/or the biofunctionalization of surfaces and structures of unique interest for a wide variety of applications, ranging from novel biomedical diagnostics to powerful new therapeutics to advanced biomaterials.

This book explores well-established and emerging conjugation strategies that are relevant for proteins used in the field of precision medicine, focusing on techniques that are suitable for antibodies, antibody-fragments such as Fabs, scFvs, or nanobodies, scaffold proteins such as FN3 or DARPin, peptides, or model proteins. Although centered on the development of bioconjugates rather than their application, most protocols also show the conjugation of the targeting vehicle to a diagnostic or therapeutic entity, with the end-product most often being an antibody-drug conjugate, an optical probe, a nanomedicine, or a radiopharmaceutical. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Bioconjugation: Methods and Protocols is an ideal guide for researchers looking toward precision medicine in order to expand the vital field of drug discovery.

Bioconjugate Techniques, 2nd Edition, is the essential guide to the modification and cross linking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or cross linking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. A one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab Step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates More than 600 figures that visually describe the complex reactions associated with the synthesis of bioconjugates Includes entirely new chapters on the latest areas in the field of bioconjugation as follows: Microparticles and nanoparticlesSilane coupling agentsDendrimers and dendronsChemoselective ligationQuantum dotsLanthanide chelatesCyanine dyesDiscrete PEG compoundsBuckyballs,fullerenes, and carbon nanotubesMass tags and isotope tagsBioconjugation in the study of protein interactions

Explores bioconjugate properties and applications of polymers, dendrimers, lipids, nanoparticles, and nanotubes Bioconjugation has enabled breakthroughs across many areas of industry and biomedicine. With its emphasis on synthesis, properties and applications, this book enables readers to understand the connection between chemistry and the biological application of bioconjugated materials. Its detailed descriptions of methods make it possible for researchers to fabricate and take full advantage of bioconjugates for a broad range of applications. Moreover, the book sets the foundation for the development of new applications, including assays, imaging, biosensors, drug delivery, and diagnostics. Chemistry of Bioconjugates features contributions from an international team of leading experts and pioneers in the field. These contributions reflect the authors’ firsthand laboratory experience as well as a thorough review of the current literature. The book’s six sections examine: General methods of bioconjugation Polymer bioconjugates Organic nanoparticle-based bioconjugates Inorganic nanomaterial bioconjugates, including metals and metal oxides Cell-based, hydrogel/microgel, and glyco-bioconjugates Characterization, physico-(bio)chemical properties, and applications of bioconjugates This comprehensive exploration of bioconjugates includes discussions of polymers, dendrimers, lipids, nanoparticles, and nanotubes. References at the end of each chapter serve as a gateway to the most important original research findings and reviews in the field. By drawing together and analyzing all the latest chemical methods and research findings on the physico-chemical and biochemical properties of bioconjugates, Chemistry of Bioconjugates sheds new light on the significance and potential of bioconjugation. The book is recommended for organic and polymer chemists, biochemists, biomaterial scientists, carbohydrate chemists, biophysicists, bioengineers, and drug and gene delivery scientists.

Bioconjugate Techniques, Third Edition, is the essential guide to the modification and cross linking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions, with details on hundreds of commercially available reagents and the use of these reagents for modifying or crosslinking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. Offers a one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab Provides step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates Features full color illustrations Includes a more extensive introduction into the vast field of bioconjugation and one of the most thorough overviews of immobilization chemistry ever presented

A cutting-edge collection of basic and state-of-the-art methods optimized for investigating the molecular biology of this class of retrovirus. These readily reproducible techniques range from methods for the isolation and detection of human retroviruses to cutting-edge methods for exploring the interplay between the viruses and the host. Here, the researcher will find up-to-date techniques for the isolation and propagation of HIV, HTLV, and foamy virus from a variety of sources. There are also assays for determining the cell tropism of HIV-1, the coreceptor usage of HIV-1, and human gene expression with HIV-1 infection by microarrays, as well as for phenotyping HIV-1 infected monocytes and examining their fitness. Highlights include the detection and quantification of HIV-1 in resting CD4+, a new cloning system for making recombinent virus, cDNA microarrays, and the determination of genetic polymorphisms in two recently identified HIV-1 co-factors that are critical for HIV-1 infection.

Biomedical Applications of Functionalized Nanomaterials: Concepts, Development and Clinical Translation presents a concise overview of the most promising nanomaterials functionalized with ligands for biomedical applications. The first section focuses on current strategies for identifying biological targets and screening of ligand to optimize anchoring to nanomaterials, providing the foundation for the remaining parts. Section Two covers specific applications of functionalized nanomaterials in therapy and diagnostics, highlighting current practice and addressing major challenges, in particular, case studies of successfully developed and marketed functionalized nanomaterials. The final section focuses on regulatory issues and clinical translation, providing a legal framework for their use in biomedicine. This book is an important reference source for worldwide drug and medical devices policymakers, biomaterials scientists and regulatory bodies. Provides an overview of the methodologies for biological target identification and ligand screening Includes case studies showing the development of functionalized nanomaterials and their biomedical applications Highlights the importance of functionalized nanomaterials for drug delivery, diagnostics and regenerative medicine applications