Mammalian Antibody Light Chain Gene Expression Vector
Recombinant antibodies have extensive applications in the fields of diagnostics and therapeutic medicine. Currently, there are about 180 monoclonal antibody products approved worldwide targeting specific proteins, such as PD-L1, HER2, IL-17R, and VEGF. The development of genetic engineering has facilitated successful production of these therapeutic antibodies, which have ideal tumor penetration behavior, short retention times, and reduced immunogenicity. These therapeutic recombinant antibodies are mainly produced in mammalian cells in vitro (e.g., CHO cells), since the post-transcriptional modification in this expression system is closest to the in vivo processes in humans.
VectorBuilder’s mammalian antibody light chain gene expression vector has been designed specifically for high-yield production of monoclonal antibody light chains. All antibodies contain heavy and light chains, both of which consist of variable (V) and constant (C) regions. In mammals, antibodies are divided into five isotypes (IgG, IgA, IgM, IgD and IgE) based on the heavy chain C region sequence. A given mammalian antibody isotype either has lambda (λ) or kappa (κ) light chains which are encoded by the light chain C region. While there is no apparent functional difference, the ratio of the two types of light chains varies from species to species. The light chain plays a role in preventing self-reactivity, and there is growing evidence that it enhances receptor activation through homotypic interactions.
In addition to this constant region, the V regions in heavy and light chains contribute to antigen recognition. Therefore, by customizing sequences encoding the light chain V region, customers may generate vectors to satisfy their personalized need for antigen-binding specificities. In the vector design process, the light chain C region may either be selected from our popular light chain C region database (containing human κ and λ2 light chains, and mouse κ, λ1, λ2 light chains) or be pasted by customers using their own sequence. Additionally, signal peptides (e.g., IL-2 sig) can be added into the vector to increase secretion of the recombinant antibody.
For further information about this vector system, please refer to the papers below.
|MAbs. 14:2014926 (2022)||Overview of antibody therapeutics|
|Nat Protoc. 13:99 (2018)||Overview of design for antibody expression vectors|
|Protein Expr Purif.118:105-12 (2016)||Description of signals which can increase the secretory protein production|
|Proc Natl Acad Sci USA. 119: e2201562119 (2022)||Applications of recombinant light chain antibody|
Our expression vector is optimized for high-efficiency transfection and high yield of monoclonal antibody light chains in mammalian cells. By customizing sequences encoding the variable light chains into the vector, customers may obtain antibodies with high-affinity to target antigens.
Technical simplicity: Delivering plasmid vectors into cells by conventional transfection is technically straight forward, and far easier than virus-based vectors which require the packaging of live viruses.
Reproducibility and scalability: The recombinant protein is harvested directly from transfected host cells. Therefore, reproducibility of different batches can be easily achieved. Moreover, the amplification of host cells allows for large-scale antibody production.
Non-integration of vector DNA: Conventional transfection of plasmid vectors is also referred to as transient transfection because the vector stays mostly as episomal DNA in cells without integration. However, plasmid DNA can integrate permanently into the host genome at a very low frequency (102 to 106 cells depending on cell type). If a drug resistance or fluorescence marker is incorporated into the plasmid, cells stably integrating the plasmid can be derived by drug selection or cell sorting after extended culture.
Optimal expression ratio of the heavy to light chain difficult to achieve: All antibodies consist of heavy chains and light chains. For successful antibody production, a precise expression ratio of the heavy to light chain is required. This can only be achieved by co-transfecting a second plasmid expressing the antibody light chain. Achieving optimal expression of both heavy chain and light chain may be difficult to control due to expression driven by different promoters in two distinct vectors.
Promoter: The promoter driving your gene of interest is placed here.
Kozak: Kozak consensus sequence. It is placed in front of the start codon of the ORF of interest because it is believed to facilitate translation initiation in eukaryotes.
IL2-sig: Signal peptide of Homo sapiens interleukin 2. It facilitates the secretion of the protein.
Light Chain Variable Region (VL): light chain variable region for antigen recognition
Light Chain Constant Region (CL): light chain constant region
SV40 late PA: Simian virus 40 late polyadenylation signal. It facilitates transcriptional termination of the upstream ORF.
Marker: A drug selection gene (such as neomycin resistance), a visually detectable gene (such as EGFP), or a dual-reporter gene (such as EGFP/Neo). This allows cells transduced with the vector to be selected and/or visualized.
pUC ori: pUC origin of replication. Plasmids carrying this origin exist in high copy numbers in E. coli.
Ampicillin: Ampicillin resistance gene. It allows the plasmid to be maintained by ampicillin selection in E. coli.