Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries.

Abstract

Conventional antibody generation usually requires active immunization with antigen immediately prior to the preparation procedure. Combinatorial antibody library technology offers the possibility of cloning a range of antibody specificities at a single point in time and then accessing these specificities at will. Here we show that human monoclonal antibody Fab fragments against a plethora of infectious agents can be readily derived from a single library. Further examination of a number of libraries shows that whenever antibody against a pathogen can be detected in the serum of the donor, then specific antibodies can be derived from the corresponding library. We describe the generation of human Fab fragments against herpes simplex virus types 1 and 2, human cytomegalovirus, varicella zoster virus, rubella, human immunodeficiency virus type 1, and respiratory syncytial virus. The antibodies are shown to be highly specific and a number are effective in neutralizing virus in vitro.

Repertoire cloning of lupus anti-DNA autoantibodies.

“…Two studies in mouse models have attempted to directly evaluate this relationship, but the results led to different conclusions regarding whether the library approach enabled recovery of Abs with the same V H and V L genes sampled by the hybridoma approach (39,40). While direct side-by-side comparisons of hybridomas and combinatorial library studies have not been feasible in humans, studies have documented that repertoire cloning approaches can isolate human Abs with Ag-binding properties identical to those of the donor's serum Abs to infectious pathogens (41,42) and autoAgs (43,44). In the current report, anti-dsDNA Abs were obtained from libraries from two lupus patients, and our sequence analyses and idiotype surveys indicate that these recovered autoAbs can use V regions with predominant H-L combinations that reiterate features of the in vivo response (discussed further below).…”

Isolation of human anti-c-erbB-2 Fabs from a lymph node-derived phage display library

“…The theoretical advantages of recombinant human antibodies over mouse monoclonals as therapeutic agents are well recognized [1][2][3][4][5]. However, the optimal source of immunoglobulin genes for the construction of these libraries remains uncertain.…”

“…Phage display libraries have been constructed from immunoglobulin genes derived from a wide variety of tissues, including peripheral blood, bone marrow and lymph nodes [2,6,7]. These tissues are frequently obtained from immunologically naive, healthy donors [5,6].…”

Phage display of intact domains at high copy number: A system based on SOC, the small outer capsid protein of bacteriophage T4

“…The SOC display system is capable of presenting up to -I O 3 copies per capsid and >IO4 copies per polyhead of V3-sized domains. Phage displaying SOC-VPI were isolated from a 1:106 mixture by two cycles of a simple biopanning procedure, indicating that proteins of at least 35 kDa may be accommodated.Keywords: bacteriophage T4; capsid assembly; human immunodeficiency virus antigens; phage display; protein engineering Filamentous phage-based display systems (Smith, 1985) have found widespread use in molecular biology, including many immunologic applications such as antigen presentation and the immunoisolation of desired recombinants (biopanning) (Marks et al, 1992;Smith et al ., 1993;Williamson et al, 1993). However, with filamentous phages, peptides that may be displayed from the major coat protein are limited in size to 6-10 amino acid residues (Kishchenko et al, 1994;Iannolo et al, 1995), although somewhat…”