ABSTRACT
Three signicant developments in molecular biology have been central in promoting the growing role of antibodies as tools for biochemical and biological research and as a signicant class of molecules for drug development. First, following the establishment of reliable methods for the generation of high-afnity, high-specicity
CONTENTS
15.1 Introduction .................................................................................................. 347 15.2 Humanization Using Phage Display ............................................................. 350
15.2.1 Choice of Human Scaffold ............................................................... 351 15.2.2 Design of Humanization Libraries ................................................... 351 15.2.3 Further Optimization of Humanized Antibodies ............................. 352
15.3 In Vitro Afnity Maturation of Antibodies .................................................. 355 15.3.1 Library Size ...................................................................................... 358 15.3.2 Targeting Positions for Random Mutagenesis .................................. 359 15.3.3 Method of Mutagenesis..................................................................... 361 15.3.4 Diversity and Codon Degeneracy ..................................................... 363 15.3.5 Antigen-Binding Selection Methods ................................................364 15.3.6 Merging Results from Separate Libraries ........................................ 365 15.3.7 Antibody Format...............................................................................366
