Antibody Methods and Protocols

This Methods in Molecular Biology(TM) volume covers in vitro and in vivo generation of antibodies, as well as techniques for screening, analysis and modification of antibodies and antibody fragments. Offers materials lists, protocols and troubleshooting tips.

The rapidly growing field of antibody research is the result of many advancing technologies allowing current developments to take advantage of molecular engineering to create tailor-made antibodies. Antibody Methods and Protocols attempts to provide insight into the generation of antibodies using in vitro and in vivo approaches, as well as technical aspects for screening, analysis, and modification of antibodies and antibody fragments. The detailed volume is focused on basic protocols for isolating antibodies and, at the same time, it selects a range of specific areas with the aim of providing guides for the overall process of antibody isolation and characterization as well as protocols for enhancing classical antibodies and antibody fragments. Written in the highly successful Methods in Molecular Biology™ series format, 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 easy to use, Antibody Methods and Protocols provides a broad and useful background to support ongoing efforts by novices and experts alike and encourages the development of new imaginative approaches to this vital area of study.

Covers general considerations about antibody generation, antibody formats, and platforms Highlights innovative approaches Features expert tips on troubleshooting and avoiding known experimental pitfalls Includes supplementary material: sn.pub/extras

Inhalt

Antigen Presentation for the Generation of Binding Molecules.- Recombinant Antibodies and In Vitro Selection Technologies.- Phage Display.- Generation of Human Fab Libraries for Phage Display.- Selection of Human Fab Libraries by Phage Display.- Ribosome Display.- Hybridoma Technology for the Generation of Monoclonal Antibodies.- The Application of Transgenic Mice for Therapeutic Antibody Discovery.- Production of Human or Humanized Antibodies in Mice.- Immortalization of Human B Cells: Analysis of B Cell Repertoire and Production of Human Monoclonal Antibodies.- Kinetic Screening in the Antibody Development Process.- Temperature-Dependent Antibody Kinetics as a Tool in Antibody Lead Selection.- Determination of Antibody Glycosylation by Mass Spectrometry.- Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format.- PEGylation of Antibody Fragments for Half-Life Extension.- Bispecific Antibody Derivatives Based on Full-Length IgG Formats.- Generation of Fluorescent IgG Fusion Proteins in Mammalian Cells.- Methods to Engineer and Identify IgG1 Variants with Improved FcRn Binding or Effector Function.- Class-Specific Effector Functions of Therapeutic Antibodies.