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Recombinant Chimeric Antibodies Customized Services

Chimeric Antibody refers to the formation of complete IgG antibody by chimerizing the variable region of antibody from one species with the constant region fragment of another species, such as human-mouse, human-rabbit, mouse-rabbit, human-alpaca chimeric antibody, etc. The main purpose of modification of chimeric antibody from different species is to improve the immunogenicity of the parental antibody. The transformation of chimeric antibodies from different species is mainly to improve the immunogenicity, species characteristics, half-life and other characteristics of the parental antibody, and the core demand is to obtain the variable region sequence of the antibody. With the help of phage display technology and yeast antibody display technology, monoclonal antibodies from various species can be discovered, and then chimeric antibodies from different sources can be obtained.

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In 2014, the WHO (World Health Organization) released new guidelines that redefined the different classes of therapeutic antibodies. Its INN (International Nonproprietary Name) nomenclature for antibody drugs classifies therapeutic antibodies into 3 categories: chimeric or "-ximab, humanized or '-zumab, and fully human or '-umab.

To classify a specific antibody according to WHO nomenclature, antibodies derived from mouse (or more generally non-human) sequences would be categorized as -zumab, while antibodies derived from humans (phage display, transgenic mice, or B-cell cloning technology) would be categorized as -umab monoclonal antibodies. In a standard program, mouse antibodies that are BLASTed against the IMGT database will typically show approximately 65-80% identity to the closest human germline in the VL structural domain and 60-75% identity in the VH structural domain. Therefore, mutation of some residues in the CDR is usually required to achieve the desired identity in the VH domain. Therefore, mutation of the CDR (as defined by Kabat) is required during humanization to achieve 85% identity in the VH structural domain.

Tek Biotech is a professional chimeric antibody construction and production company, with a mature recombinant antibody lactation expression platform, can provide customers with high-quality chimeric antibody expression customized services. The complete Gibco serum-free mammalian cell culture system, together with 293F and CHO suspension cell lines, can meet the needs of different customers for recombinant expression of natural conformational proteins in vitro. Tek Biotech provides customers with a full set of cost-effective research services to meet GMP-like system, including cell seed bank source documentation system, cell seed batch verification, quality control of animal source components, standardized cell culture and transformation operation procedures and so on.

█ Classification of Chimeric Antibodies

Chimeric Antibody Types

Synopsis

Chimeric Principle (physics)

Chimeric IgG Antibody

During immunization, the Fc segment of the antibody is able to interact with effector cells, thereby exerting a specific biological function; IgG1 and IgG3 have the strongest binding capacity to the Fc receptor, followed by IgG4, and IgG2 has almost undetectable binding.

The construction of chimeric antibodies requires firstly obtaining the V region gene from the hybridoma cells producing the murine-derived monoclonal antibody, then recombining it with the human C region gene and cloning it into a suitable vector, and then transferring it to the recipient cells for expression.

Chimeric Fab and F(ab')2

Antibodies

The biggest advantage of this type of antibody is its high penetration power. Due to the low affinity and small molecular weight of this type of antibody, it is easily filtered by the glomeruli and disappears from the bloodstream, therefore, most of them are not suitable for clinical treatment alone.

The genes of the V region of the L, H chain of the functional antibody were recombined with human light chain K and H before being transferred to host cells for expression.

█ Strategies for Constructing Human-mouse Chimeric Antibodies

1. Template replacement: Human antibody FR with greater homology to the murine counterpart is used to replace the murine FR, using variable region frameworks (VH such as NEW, KOL, VL such as REI, etc.) of human antibodies with existing crystal structure data as the basic template for replacement, or searching for a human FR with maximum homology to the murine McAb FR in the existing antibody sequence libraries for replacement.

2. Surface remodeling: veneering or resurfacing of the surface residues of murine CDR and FR to make them resemble the profile of human CDR or human FR.

3. Compensatory transformation: Select residues in the human FR that interact with the CDR, have a close relationship with the affinity of the antibody, or play a key role in the folding of the FR spatial structure, and change them to compensate for the complete CDR transplantation.

4. Localization retention: Humanized McAb retains amino acid residues involved in antigen binding in the variable region of murine McAb, including some key residues in CDR and FR.

█ Chimeric Antibody Design and Expression Service Cycle

Step	Service Content	Period
Gene cloning and vector construction	* Amplification/identification of gene fragments or synthesis of target genes for insertion into mammalian cell expression plasmids * PCR and sequencing assays to identify subcloned genes * Delivery: sequencing data and experimental protocol	1-2 weeks
Transient protein expression	* Recombinant plasmid transfected cells (293F, CHO): transient expression * SDS-PAGE and WB detection * Delivery: Expression vector, cloned strain, recombinant antibody, purity >85	2–3 weeks
Stabilization of expression (optional)	* Stable expression cell line construction and screening of recombinant protein CHO * Delivery: stable cell line, construction report; marginal product, 3 clones, 50 ml fermentation product and purification product per clone	12–14 weeks

█ Service Advantage

-- Variable region multiple genus selection: human, mouse, dog, chicken, camel, etc.

-- Free recombinant chimeric antibody expression program design

-- Original optimized HEK293/CHO platform for better glycosylation and post-translational modifications

-- Self-designed expression vectors for mammalian systems

-- Over-sized and amplified expression systems are available.

-- Multiple genera available in the constant region: human, rat/mouse, canine, rhesus monkey, rabbit, etc.

-- Experience with multi-subtype constructs: IgG, IgA, IgM and IgE

Recombinant Chimeric Antibodies Customized Services Frequently Asked Questions

Q: The expression level of recombinant chimeric antibodies in the host system is low, resulting in insufficient production.

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.
Q: The structure or activity of recombinant chimeric antibodies is affected, resulting in incomplete function or inability to express them normally.

A: Optimize the design of antibody structure to ensure its correct conformation and function; Evaluate the activity and stability of chimeric antibodies through structural prediction and analysis; Perform functional validation to confirm the activity and specificity of the antibody.
Q: The selection and design of antigens may affect the binding efficiency and specificity of recombinant chimeric antibodies.

A: Optimize the design and expression of antigens to ensure their binding efficiency and specificity with recombinant chimeric antibodies; 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.
Q: The purification and stability of recombinant chimeric antibodies may be affected, affecting the reliability of subsequent applications.

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.

Q: The expression level of recombinant chimeric antibodies in the host system is low, resulting in insufficient production.

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.
Q: The structure or activity of recombinant chimeric antibodies is affected, resulting in incomplete function or inability to express them normally.

A: Optimize the design of antibody structure to ensure its correct conformation and function; Evaluate the activity and stability of chimeric antibodies through structural prediction and analysis; Perform functional validation to confirm the activity and specificity of the antibody.
Q: The selection and design of antigens may affect the binding efficiency and specificity of recombinant chimeric antibodies.

A: Optimize the design and expression of antigens to ensure their binding efficiency and specificity with recombinant chimeric antibodies; 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.
Q: The purification and stability of recombinant chimeric antibodies may be affected, affecting the reliability of subsequent applications.

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.

Consult Now Recombinant Chimeric Antibodies Customized Services

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Recombinant Chimeric Antibodies Customized Services Frequently Asked Questions

Q: The expression level of recombinant chimeric antibodies in the host system is low, resulting in insufficient production.

Q: The structure or activity of recombinant chimeric antibodies is affected, resulting in incomplete function or inability to express them normally.

Q: The selection and design of antigens may affect the binding efficiency and specificity of recombinant chimeric antibodies.

Q: The purification and stability of recombinant chimeric antibodies may be affected, affecting the reliability of subsequent applications.

Q: The expression level of recombinant chimeric antibodies in the host system is low, resulting in insufficient production.

Q: The structure or activity of recombinant chimeric antibodies is affected, resulting in incomplete function or inability to express them normally.

Q: The selection and design of antigens may affect the binding efficiency and specificity of recombinant chimeric antibodies.

Q: The purification and stability of recombinant chimeric antibodies may be affected, affecting the reliability of subsequent applications.

Consult Now Recombinant Chimeric Antibodies Customized Services