Baculovirus cassette vector pAc-k-Fc for the expression of human, humanized or chimeric IgG(kappa) 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 Fab 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 genefragments 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 reportedpreviously (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. Now available are 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 cloning of heavy and light chain Fv or Fab gene fragments isolated from hybridomas, individual B cell clones as well as antibody libraries.
Expression vector pAc-κ-Fc
Vector name pAc-κ-Fc
Category Baculovirus cassette vector for the expression of human,humanized or chimeric IgG(κ) in insect cells
Application Cloning of heavy and light chain genes of Fab antibody fragments selected from hybridomas, single B-lymphocytes or phage display libraries
Cloning sites: XhoI and SpeI for heavy chain Fab fd genes SacI and EcoRV for light chain Fab(κ) genes. Expression of immunoglobulin heavy and light chains in insectcells and secretion of assembled IgG(κ) antibodies into the supernatant Vector size 6702 bp Resistance The vector contains an Ampicillin resistance marker gene.
Vector Size:6702bp
Resistance:Vector contains an ampicillin resistance marker gene
Applications:Cloning of heavy and light chain variable domain genes of scFv antibody fragments selected from hybridomas, single B-lymphocytes or phage display libraries.
Cloning sites:XhoI and SpeI for heavy chain Fab fd genes SacI and EcoRV for light chain Fab(k) genes. Expression of immunoglobulin heavy and light chains in insect cells and secretion of assembled IgG(k) antibodies into the supernatant .
Storage and Stability:For long-term storage, store at -20°C. Aliquots are stable for at least 6 months at -20°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.
