Baculovirus Expression Vector pAc-l-CH3

Baculovirus cassette vector pAc-l-CH3 for the expression of human, humanized or chimeric IgG(lambda) in insect cells and secretion of assembled antibodies into the supernatant. The vector derived from pAcUW51 is suitable for cloning of heavy and light chain scFv antibody gene fragments. Within the past decade, recombinant antibody technologies have been widely used to produce various single-chain Fv or Fab antibody fragments of different specificity. The randomized combination of cloned variable heavy and light chain immunoglobulin gene fragments further allowed the construction of human antibody libraries, which enable today the isolation of specific scFv or Fab antibody fragments against particular antigens e.g. by phage display. For many applications, however, it is required to reassemble the variable regions of the selected antibody with immunoglobulin constant regions to generate complete antibody molecules. The baculovirus expression system has already been established as a reliable system for the production of immunoglobulins (Nesbit et al., 1992; Liang et al., 1997, 2001). Even the development of cassette vectors for the production of human antibodies have been reported previously (Poul et al., 1995a,b). However, these systems are based on a combination of two different vectors which separately served for the expression of light and heavy chains. The required careful and time consuming adjustment of the two respective recombinant baculovirus titers now becomes obsolete, since we offer new single baculovirus cassette vectors with authentic IgG heavy and light chain signal sequences for the rapid production of complete chimeric, humanized and human IgG antibodies in recombinant baculovirus infected insect cells (Liang et al., 2001). The vectors were specifically designed for clon- ing of heavy and light chain Fv or Fab gene fragments isolated from hybridomas, individual B cell clones as well as antibody libraries. Vector Size: 7360bp Resistance: Vector contains an ampicillin resistance marker gene Applications: Cloning of heavy and light chain variable domaingenes of scFv antibody fragments selected from hybridomas, single B-lymphocytes or phage display libraries. Cloning sites: Xhol and Nhel for heavy chain VH genes. Sacl and HindIII for light chain VL(k) genes. Storage and Stability: May be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.