MAKING MONOCLONAL-ANTIBODIES BY REPERTOIRE CLONING

In this review we will focus on the applications of the basic knowledg e gained by analysing immunoglobulin gene sequences and their transcri ption and translation. This knowledge allowed the isolation of immunog lobulin genes from a polyclonal source of lymophocytes by the polymera se chain reaction. In parallel, filamentous bacterial phage technology was adapted to express immunoglobulin genes in Escherichia coli in or der to produce phage libraries containing 10(7-11) different immunoglo bulin genes. The possibility of cloning such a huge repertoire allowed to immunize animals with a mixture of antigens followed by the isolat ion of the desired monoclonal antibody from the phage library. Future work will aim at in vitro manipulating genes encoding immunoglobulins to obtain antibodies with desired specificity and biological function.