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Recombinant IgG Customization Services
Recombinant monoclonal antibody is an antibody produced by recombinant DNA technology, specifically, it refers to the DNA recombination technology to modify and recombine the corresponding gene sequences of the antibody according to the needs, and constructed on a plasmid, and then through the exogenous protein expression technology, the constructed plasmid is transfected/transformed into the appropriate host cell to express the antibody. Recombinant antibodies are a good solution to the human rejection caused by animal-derived antibodies, making antibodies humanized and more effective.
Tek Biotech has a complete recombinant antibody expression production platform, rich experience in recombinant protein and recombinant antibody expression, and is able to provide customers with recombinant IgG antibody expression services for various species and genera, including human, mouse, rabbit, sheep, dog, camel, cow, etc. Customers only need to provide the antibody sequence information, hybridoma cells, or cDNA, and the technical support of the company can carry out the design and optimization of the expression system according to the customer's needs. The technical support of the company can design the scheme and optimize the expression system according to the customer's needs, combined with the company's independently designed breastfeeding system of high-expression vectors and large-scale recombinant antibody fermentation platform, to provide customers with high-quality recombinant antibody preparation services.
Recombinant antibody development methods are mainly classified into three categories: 1. Phage display technology: High-affinity peptides that bind to specific receptors can be screened from large-capacity antibody peptide libraries through phage display technology. Combined with recombinant technology, the target gene can be cloned and the expressed antibody protein can be displayed and screened for antibodies with high specificity and affinity by phage display technology. Tek Biotech can provide recombinant antibody production based on phage display technology, please contact us for details. 2. B-cell cloning technology: plasma cells are isolated through animal immunization and separated by flow cytometry. The cellular genes are amplified and screened, cloned into vectors, and the better expressed clones are screened using ELISA and sequenced, so that recombinant antibodies can be prepared in vitro. 3. Hybridoma sequencing: gene sequencing is performed on the prepared hybridomas into the antibody obtained, and the antibody is expressed using recombinant technology in vitro.
█ Recombinant Antibody Preparation Process
█ IgG Immunization Effect
-- Fab segment binds to antigen and exerts a neutralizing effect;
-- Complement-dependent cytotoxicity (CDC): the Fc segment activates complement and forms membrane attack complexes capable of lysing target cells;
-- Conditioning and phagocytosis (ADCP): the Fc segment can bind to Fc receptors on the surface of neutrophils and macrophages and enhance phagocytosis;
-- Antibody-dependent cell-mediated cytotoxicity (ADCC): the Fc segment binds to the Fc receptor on the surface of natural killer cells and mediates the killing of target cells by natural killer cells.
█ Antibody Type
Five classes of antibodies have been found in human serum, as shown below:
IgA | IgD | IgE | IgG | IgM | |
Heavy chain | α | δ | ε | γ | μ |
Placement | Intravascular and secretion | B-cell surface | Basophils and mast cells in saliva and nasal secretions | Endovascular and extravascular | Mainly in blood vessels |
Function | Protecting the mucus membrane | Currently unknown | Prevents parasites and is associated with allergic reactions | Secondary immune response | Primary immune response |
Subtype | IgA1 IgA2 | 1 | 1 | IgG1、IgG2、IgG3、IgG4 | 1 |
Fc region binds to | Bone marrow cells | B-cell | Mast cells and basophils | Phagocyte (cell that ingests and destroys foreign matter) | B T and NK cells |
IgD、IgE and IgG are usually found as monomers, while IgA is usually found as a dimer and IgM as a pentamer.
█ Antibodies to Different IgG Subtypes
Species | IgG type |
Human | IgG1/2/3/4 |
Mouse | IgG1/2a/2b/2c/3 |
Rat | IgG1/2a/2b/2c |
Rabbit | IgG |
Dog | IgG |
Tek Biotech can provide recombinant antibody preparation services for all of the above species and isoforms.
█ Advantages of Recombinant Antibody Preparation
-- The structure of the antibody can be modified and optimized in a series of ways (including the introduction of humanization, the Fc region of the human IgG1 class, etc.) to improve the affinity and stability of the antibody.
-- Wider selection of antigens, not limited by source, purity and quantity.
-- No need to immunize animals, reducing the number of animal experiments and ethical issues.
-- The resulting antibodies have greater specificity and affinity and can be used in therapeutic and detection areas.
█ Services for Recombinant Antibody Preparation
Process | Service Content | Delivery |
Gene synthesis and codon optimization | Customers can provide antibodies/amino acid sequences or recombinant plasmids containing antibody genes | 1.purify antibodies 2. Expression plasmids 3. QC report |
Vector construction | Cloning into Tektronix High Efficiency Expression Vectors; Plasmid Extraction | |
Expression purification | Transient transfection of HEK293/CHO cells; Purification of antibodies | |
Quality control analysis | SDS-PAGE; UV; endotoxin analysis, ELISA, etc. (optional) |
█ Service Advantages
-- Full-length and scFv, Fab, VHH, chimeric antibodies, bispecific antibodies, Fc fusion antibodies from multiple species sources (human, mouse, rabbit, etc.) have been successfully expressed.
-- Mammalian cell expression platform: self-designed high-expression vectors, optimized proprietary culture media and proprietary transfection reagents.
-- Multiple specifications of amplified expression systems to meet different customer demand volumes.
-- Professional technical team to customize personalized one-to-one antibody discovery total solutions.
A: Optimize the construction of expression vectors, including selecting appropriate promoters, signal sequences, and host systems; Optimize cultivation conditions, such as temperature, medium composition, oxygen supply, etc., to increase expression levels; Consider using enhancers or expression adjuvants to improve the expression efficiency of antibodies.
A: Optimize the structure of IgG to ensure its correct conformation and function; Evaluate the activity and stability of IgG through structural prediction and analysis; Perform functional validation to confirm the activity and specificity of IgG.
A: Optimize the design and expression of antigens to ensure their binding efficiency and specificity with recombinant IgG; Conduct preliminary antigen assessment and screening to select the most suitable antigen; Consider using affinity modification or structural optimization methods to improve the binding efficiency and specificity between antibodies and antigens.
A: Optimize purification methods and steps, select appropriate purification columns, chromatography conditions, and elution buffer to improve purification efficiency; Conduct stability testing to evaluate the stability and storage conditions of antibodies; Consider using additives or protectants to improve the stability and shelf life of antibodies.
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