On the use of combinatorial antibody libraries to clone the "fossil record" of an individual's immune response

For about the last century the record of immunological events could only be obtained from serum proteins. We suggest that in the future this will change and the far more detailed nucleic acid record will provide new insights into immunological processes as well as selective access to the complete repertoire

Linkage of recognition and replication functions by assembling combinatorial antibody Fab libraries along phage surfaces

We describe a method based on a phagemid vector with helper phage rescue for the construction and rapid analysis of combinatorial antibody Fab libraries. This approach should allow the generation and selection of many monoclonal antibodies. Antibody genes are expressed in concert with phage morphogenesis, thereby allowing incorporation of functional Fab molecules along the surface of ifiamentous phage. The power of the method depends upon the linkage of recognition and replication functions and is not limited to antibody molecules

In vitro selection and affinity maturation of antibodies from a naive combinatorial immunoglobulin library

We have used a combinatorial immunoglobulin library approach to obtain monoclonal antibodies from nonimmune adult mice, thereby establishing the principles of (i) accessing naive combinatorial antibody libraries for predetermined specificities and (ii) increasing the affinity of the selected antibody binding sites by random mutagenesis. A combinatorial Fab library expressing immunoglobulin Ip and it light-chain fragments on the surface of filamentous phage was prepared from bone marrow of nonimmunized, adult BALB/c mice with the multivalent display vector pComb8. Phage displaying low affinity Fabs (binding constants, 104-1O M-') binding to a progesterone-bovine serum albumin conjugate were isolated from the library. Random mutagenesis of the heavy- and light-chain variable regions expressed in the monovalent phage display vector pComb3 was performed by errorprone PCR, and subsequently clones with improved affinity for the hapten conjugate were selected. We demonstrate that antibodies with desirable characteristics from a nonimmune source may be selected and affinity maturation may be achieved by using the twin vectors pComb8 and pComb3, thus opening the route to obtaining specific antibodies from a generic library and bypassing immunization