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Recombinant Chimeric Antibodies Customized Services

TekBiotech focuses on developing high-quality targeted drug antibodies for customers. In order to help customers conduct high-quality drugability evaluation, we have built an in vitro recombinant expression platform with mammalian cells as the core, which can provide customers with high-quality one-stop recombinant antibody preparation services including sequence analysis, linker design, fusion tag design, high expression vector design and construction, expression scheme design optimization, recombinant expression condition optimization, antibody purification, etc.

Chimeric antibody refers to a complete IgG antibody formed by chimerizing the variable region of an antibody from one species with the constant region fragment of another species, such as human-mouse, human-rabbit, mouse-rabbit, human-alpaca chimeric antibodies, etc., as shown in Figure 1. The modification of chimeric antibodies of different species is mainly to improve the immunogenicity, species characteristics, half-life and other characteristics of the parent antibody. Its core appeal is to obtain the variable region sequence of the antibody. With the help of phage display technology and yeast antibody display technology, monoclonal antibodies of various species can be discovered, and chimeric antibodies from different sources can be obtained.

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Figure 1 Schematic diagram of human-mouse chimeric antibodies

█ Types of Chimeric Antibody Expression Services

Using our recombinant expression technology platform, TekBiotech can provide customers with the following related chimeric antibody recombinant expression services, as shown in Table 1.

Table1 Types of recombinant antibody expression services

Option 1: Chimeric complete IgG antibody

Option 2: Chimeric Fab and F(ab')2 antibodies

█ Chimeric Recombinant Antibody Expression Services

TekBiotech is a professional chimeric antibody construction and production company with a mature recombinant antibody mammalian expression platform, which can provide customers with high-quality chimeric antibody expression customization services. A full set of Gibco serum-free mammalian cell culture, combined with 293F and CHO suspension cell lines with clear sources, the entire system meets the in vitro recombinant expression needs of different customers for natural conformation proteins. The full set of cost-effective research services provided by TekBiotech to customers meet the GMP-like system, including the cell seed bank source document record system, cell seed batch verification, animal-derived component quality control, standardized cell culture and transformation operation procedures, etc.

Table2 Contents of chimeric recombinant antibody expression service

Steps	Service Content	Period
Step1: Gene cloning and vector construction	-- Amplify/identify gene fragments or synthesize target genes, and insert them into mammalian cell expression plasmids; -- PCR and sequencing tests to determine subcloned genes; -- Delivery: sequencing raw data and experimental report;	1-2 Weeks
Step2: Transient protein expression	-- Recombinant plasmid transfected cells (293F, CHO) transient expression + purification; -- SDS-PAGE and WB detection; -- Delivery: expression vector, cloning strain, recombinant antibody, purity>85%, experimental report;	1-2 Weeks
Step3: Stable expression (optional)	-- Construction and screening of recombinant protein CHO stable expression cell line; -- Delivery: stable cell line, construction report; marginal product, 3 clones, 50ml fermentation product and purified product for each clone;	12–14 Weeks

Tekbiotech can also provide customers with one-stop technical services such as antibody humanization, CAR-T/CAR-NK lead sequence design and cell killing verification, to help customers' scientific research projects and drug antibody development.

█ Advantages of Antibody Recombinant Expression Service Platform

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Fast cycle, high success rate

Proprietary expression vector system,

efficient expression

Strict QC control system

Guaranteed high-quality service

Recombinant Chimeric Antibodies Customized Services Frequently Asked Questions

What are recombinant chimeric antibodies and why are they used?

Recombinant chimeric antibodies are antibodies that are made up of antibody domains from different species (usually human and mouse). The basic structure is to recombinant the antigen binding site of a mouse antibody with the constant region of a human antibody to construct a partially humanized antibody. The purpose of this is to maintain the original specificity and affinity of the mouse antibody while reducing the immune response caused by the mouse antibody. The advantage of recombinant chimeric antibodies is that they have high antigen specificity while avoiding the immune rejection of fully mouse antibodies in the human body, especially in the development of therapeutic antibodies, such antibodies can improve efficacy and reduce side effects. This type of antibody is widely used in the diagnosis and treatment of diseases and in the field of biopharmaceuticals, especially in cancer immunotherapy.
What are the advantages and disadvantages of recombinant chimeric antibodies compared to fully human antibodies?

Recombinant chimeric antibodies have certain advantages and disadvantages compared to fully human antibodies. In terms of advantages, recombinant chimeric antibodies can better avoid the rejection of the immune system in the human body because they contain human constant regions, while retaining the high specificity and affinity of mouse antibodies. Compared with fully human antibodies, chimeric antibodies have a shorter development cycle and lower costs because they can obtain the desired antibodies more quickly through recombinant technology, and most of the specificity has been obtained through mouse immunization. Therefore, chimeric antibodies have certain advantages in antibody screening and early drug development. On the disadvantage side, although chimeric antibodies reduce the constant region of mouse origin, they may still induce a mild immune response. Compared with fully human antibodies, chimeric antibodies have a slightly higher immunogenicity, so adverse reactions or immune tolerance problems may occur in clinical applications. Fully human antibodies can better avoid this immune rejection problem and generally have lower immunogenicity in clinical treatment.
How to optimize the affinity and specificity of recombinant chimeric antibodies?

Optimizing the affinity and specificity of recombinant chimeric antibodies is a key step in the antibody customization process. First, antibodies with high affinity can be screened through antibody affinity maturation technology (such as recombinant phage display technology or yeast display technology). This technology can gradually optimize the binding strength of antibodies to antigens through multiple rounds of screening to ensure that the final antibodies have the best affinity. Secondly, specificity optimization can be carried out through the modification of antibody sequences, such as mutation of the variable region of the antibody, site-directed mutation, etc., so as to improve the targeting ability of the antibody and reduce cross-reaction with non-target antigens. Genetic engineering methods, such as antibody library screening or humanization technology, can also help enhance the specificity of antibodies and reduce immunogenicity. Through these technical means, the optimized recombinant chimeric antibodies can show stronger efficacy and fewer side effects in clinical treatment.
What are the technical challenges in the production and purification of recombinant chimeric antibodies?

There are many technical challenges in the production and purification of recombinant chimeric antibodies. First, the selection of host cells is crucial for the expression and folding of antibodies. CHO cells, HEK293 cells and other mammalian cells are usually commonly used host cells because they can produce antibodies with correct folding and glycosylation modifications. However, host cells may face problems such as low expression and unstable yield, and it is necessary to optimize culture conditions and select a suitable expression system. Secondly, the difficulty in the purification process lies in how to efficiently separate recombinant antibodies. Common purification techniques include affinity chromatography, ion exchange chromatography, and gel filtration, but due to the heterogeneity of antibodies, multiple rounds of purification are often required to obtain high-purity antibodies. In addition, the stability of antibodies is also one of the challenges in the production process, especially for antibodies that are stored for a long time or produced on a large scale, and appropriate stability testing and optimization measures need to be taken.
What are the common quality control standards for customized recombinant chimeric antibody services?

The quality control standards for customized recombinant chimeric antibody services involve multiple aspects to ensure the effectiveness, safety, and availability of antibodies. First, the affinity and specificity of antibodies are the core indicators in quality control, which are usually tested by ELISA, surface plasmon resonance (SPR) and other technologies. Secondly, the purity of antibodies needs to be analyzed by SDS-PAGE, Western Blot, HPLC and other methods to ensure that the impurity content of antibodies is within an acceptable range. In addition, the stability of antibodies also needs to be tested, including storage stability at different temperatures, freeze-thaw stability, etc. Immunogenicity assessment is another important quality control link, and the immune response of antibodies is tested by mouse or monkey models. In addition, for therapeutic antibodies, it is also necessary to evaluate their biological activities, such as cytotoxicity, antigen binding ability, etc. A number of quality control standards are integrated to ensure that customized antibodies have excellent performance and potential for clinical application.

What are recombinant chimeric antibodies and why are they used?

Recombinant chimeric antibodies are antibodies that are made up of antibody domains from different species (usually human and mouse). The basic structure is to recombinant the antigen binding site of a mouse antibody with the constant region of a human antibody to construct a partially humanized antibody. The purpose of this is to maintain the original specificity and affinity of the mouse antibody while reducing the immune response caused by the mouse antibody. The advantage of recombinant chimeric antibodies is that they have high antigen specificity while avoiding the immune rejection of fully mouse antibodies in the human body, especially in the development of therapeutic antibodies, such antibodies can improve efficacy and reduce side effects. This type of antibody is widely used in the diagnosis and treatment of diseases and in the field of biopharmaceuticals, especially in cancer immunotherapy.
What are the advantages and disadvantages of recombinant chimeric antibodies compared to fully human antibodies?

Recombinant chimeric antibodies have certain advantages and disadvantages compared to fully human antibodies. In terms of advantages, recombinant chimeric antibodies can better avoid the rejection of the immune system in the human body because they contain human constant regions, while retaining the high specificity and affinity of mouse antibodies. Compared with fully human antibodies, chimeric antibodies have a shorter development cycle and lower costs because they can obtain the desired antibodies more quickly through recombinant technology, and most of the specificity has been obtained through mouse immunization. Therefore, chimeric antibodies have certain advantages in antibody screening and early drug development. On the disadvantage side, although chimeric antibodies reduce the constant region of mouse origin, they may still induce a mild immune response. Compared with fully human antibodies, chimeric antibodies have a slightly higher immunogenicity, so adverse reactions or immune tolerance problems may occur in clinical applications. Fully human antibodies can better avoid this immune rejection problem and generally have lower immunogenicity in clinical treatment.
How to optimize the affinity and specificity of recombinant chimeric antibodies?

Optimizing the affinity and specificity of recombinant chimeric antibodies is a key step in the antibody customization process. First, antibodies with high affinity can be screened through antibody affinity maturation technology (such as recombinant phage display technology or yeast display technology). This technology can gradually optimize the binding strength of antibodies to antigens through multiple rounds of screening to ensure that the final antibodies have the best affinity. Secondly, specificity optimization can be carried out through the modification of antibody sequences, such as mutation of the variable region of the antibody, site-directed mutation, etc., so as to improve the targeting ability of the antibody and reduce cross-reaction with non-target antigens. Genetic engineering methods, such as antibody library screening or humanization technology, can also help enhance the specificity of antibodies and reduce immunogenicity. Through these technical means, the optimized recombinant chimeric antibodies can show stronger efficacy and fewer side effects in clinical treatment.
What are the technical challenges in the production and purification of recombinant chimeric antibodies?

There are many technical challenges in the production and purification of recombinant chimeric antibodies. First, the selection of host cells is crucial for the expression and folding of antibodies. CHO cells, HEK293 cells and other mammalian cells are usually commonly used host cells because they can produce antibodies with correct folding and glycosylation modifications. However, host cells may face problems such as low expression and unstable yield, and it is necessary to optimize culture conditions and select a suitable expression system. Secondly, the difficulty in the purification process lies in how to efficiently separate recombinant antibodies. Common purification techniques include affinity chromatography, ion exchange chromatography, and gel filtration, but due to the heterogeneity of antibodies, multiple rounds of purification are often required to obtain high-purity antibodies. In addition, the stability of antibodies is also one of the challenges in the production process, especially for antibodies that are stored for a long time or produced on a large scale, and appropriate stability testing and optimization measures need to be taken.
What are the common quality control standards for customized recombinant chimeric antibody services?

The quality control standards for customized recombinant chimeric antibody services involve multiple aspects to ensure the effectiveness, safety, and availability of antibodies. First, the affinity and specificity of antibodies are the core indicators in quality control, which are usually tested by ELISA, surface plasmon resonance (SPR) and other technologies. Secondly, the purity of antibodies needs to be analyzed by SDS-PAGE, Western Blot, HPLC and other methods to ensure that the impurity content of antibodies is within an acceptable range. In addition, the stability of antibodies also needs to be tested, including storage stability at different temperatures, freeze-thaw stability, etc. Immunogenicity assessment is another important quality control link, and the immune response of antibodies is tested by mouse or monkey models. In addition, for therapeutic antibodies, it is also necessary to evaluate their biological activities, such as cytotoxicity, antigen binding ability, etc. A number of quality control standards are integrated to ensure that customized antibodies have excellent performance and potential for clinical application.

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Recombinant Chimeric Antibodies Customized Services Frequently Asked Questions

What are recombinant chimeric antibodies and why are they used?

What are the advantages and disadvantages of recombinant chimeric antibodies compared to fully human antibodies?

How to optimize the affinity and specificity of recombinant chimeric antibodies?