In vivo versus in vitro screening or selection for catalytic activity in enzymes and abzymes.
The recent development of catalytic antibodies and the introduction of new techniques to generate huge libraries of random mutants of existing enzymes have created the need for powerful tools for finding in large populations of cells those producing the catalytically most active proteins. Several approaches have been developed and used to reach this goal. The screening techniques aim at easily detecting the clones producing active enzymes or abzymes; the selection techniques are designed to extract these clones from mixtures. These techniques have been applied both in vivo and in vitro. This review describes the advantages and limitations of the various methods in terms of ease of use, sensitivity, and convenience for handling large libraries. Examples are analyzed and tentative rules proposed. These techniques prove to be quite powerful to study the relationship between structure and function and to alter the properties of enzymes.
Fastrez
J
J
Laboratoire de Biochimie Physique et des Biopolymères, Louvain-la-Neuve, Belgium.
D017151:Antibodies, Catalytic / Q000032:analysis* / Q000235:genetics; D002384:Catalysis; D004795:Enzyme Stability; D004798:Enzymes / Q000032:analysis* / Q000235:genetics; D019143:Evolution, Molecular; D015723:Gene Library; D006801:Humans; D016337:RNA, Catalytic / Q000378:metabolism
D017151:Antibodies, Catalytic; D004798:Enzymes; D016337:RNA, Catalytic