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Recombinant Fragment Antibody Customization Service

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.

Antigen binding fragment (Fab) is the region on the antibody that binds to the antigen. It contains a constant region and a variable region for each heavy chain and light chain. Only the variable regions of the heavy and light chains are fused together to form a single-chain variable fragment (scFv), which is only half the size of the Fab fragment, but retains the original specificity. Another small antibody fragment, single-domain antibody, also known as domain antibody, VHH, VNAR or sdAb, is an antibody fragment composed of a single monomer variable antibody domain, lacking the conventional light and heavy chain CH domains.

TekBiotech is a professional CRO service provider that can provide our customers with high-quality recombinant antibody expression services, such as scFv antibody fragments, Fab antibody fragments, sdAb single domain antibodies, heavy chain antibodies and other recombinant antibody construction and expression services, as shown in Figure 1. In addition, TekBiotech's 5,000 square meter GMP research center currently under construction can provide batch recombinant antibody production services for various industrial customers.

Schematic diagram of fragment antibodies-tekbiotech1.png

Figure1 Schematic diagram of fragment antibodies

█ Types and Characteristics of Fragment Antibodies

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

Table 1 Fragment antibody type

Antibody Type	Component	Molecular Weight (kDa)	Valence	Half-life (h)
Fab	VH+CH1+VL+CL (Light and heavy chains are connected by disulfide bonds)	50-55	Monomer	12-20
scFv	VH+VL	30	Monomer	2-4
sdAb	VHH	15	Monomer	0.5
Fc fusion antibody	(Antibody+CH2+CH3)*2	/	Dimer	/
di-scFv	scFv*2	60	Dimer	4-8

█ Common Hosts for Antibody Fragment Expression Systems

Expression System	Strain/cell	Features
Prokaryotic bacteria (E. coli)	BL21	Applicable to VHH antibodies, which will affect affinity
Mammalian Cells	CHO-S	It is a commonly used cell line in the laboratory. It grows well as an adherent and suspension culture, has high productivity, and is ideal for large-scale culture and GMP procedures; it can undergo many post-translational modifications to obtain biosimilars, showing excellent biocompatibility and drug activity; it is suitable for protein-free, animal-free production, serum-free culture conditions, and has better stability and safety.
	CHO-K1
	Freestyle 293F	It is a wild-type 293 cell line that can express proteins at high levels in serum-free conditions, and can express proteins efficiently.
	Expi 293F	It is highly transfectable, shows high protein expression levels, and exhibits faster cell growth and higher culture survival rates than standard suspension-adapted 293 cells.

█ Fragment Recombinant Antibody Expression Service

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.

Table 2 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

Note: Fab type antibodies are more difficult to express, and the project cycle will increase according to actual conditions

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

Detailed experimental design

and demonstration

Low endotoxin level:

LAL assay<0.1EU/ug

Multiple expression modes: stable expression

and transient expression

Guaranteed high-quality service: Complete

document traceability system to ensure that

all materials, reagents and preparation

information of your products are traceable

Recombinant Fragment Antibody Customization Service Frequently Asked Questions

What are recombinant fragment antibodies and why should we choose this form of antibody?

Recombinant fragment antibodies refer to recombinant antibodies composed of antibody fragments (such as Fab, scFv, Fv, etc.). Compared with traditional complete antibodies (IgG), recombinant fragment antibodies remove the constant region of the antibody and retain only the variable region, thus having a smaller molecular weight. This gives recombinant fragment antibodies some unique advantages, such as better tissue permeability, lower immunogenicity, and faster molecular clearance rate. In targeted therapy and diagnostic applications, recombinant fragment antibodies can more effectively enter tumor cells or other target tissues, so they are widely used in tumor immunotherapy, vaccine development, pathological detection, etc. Fragment antibodies produced by recombinant technology can achieve higher specificity and affinity, thus showing higher advantages in research and clinical applications.
What are the main differences between recombinant fragment antibodies and full-length antibodies?

The main difference between recombinant fragment antibodies and full-length antibodies lies in their structure and function. Full-length antibodies are usually composed of two heavy chains and two light chains, including variable regions and constant regions. The constant region plays an important role in the immune function of antibodies (such as ADCC, CDC, etc.), while fragment antibodies only retain the variable region of antibodies, that is, the antigen-binding fragment. This makes fragment antibodies small in size and simple in structure, and usually have no immune effector function, such as no activation of antibody-dependent cell-mediated cytotoxicity (ADCC) and classical complement-dependent cytotoxicity (CDC). The advantage of fragment antibodies is that they can better penetrate into tissues or cells, have faster serum clearance, and are generally less immunogenic than full-length antibodies. In addition, fragment antibodies are suitable for precision medicine and targeted therapy, such as cancer targeted drugs and molecular probes, while full-length antibodies are more used in treatments that require immune effects, such as the treatment of infections and autoimmune diseases.
What are the advantages of recombinant fragment antibody customization services?

Recombinant fragment antibody customization services have multiple advantages over traditional antibody preparation methods. First, fragment antibodies are smaller than full-length antibodies, have lower molecular weight, and have better tissue permeability, which is very important for applications such as tumor targeted therapy that require rapid penetration into cells or tissues. Secondly, fragment antibodies have reduced immunogenicity due to the removal of constant regions, so they may lead to fewer immune responses in clinical applications. In addition, the production of fragment antibodies is usually simpler and more efficient than full-length antibodies, which can reduce costs and time. Through genetic engineering technology, fragment antibodies can be optimized to improve their affinity, specificity and other properties, so as to customize antibodies that better meet the needs of customers. Finally, recombinant fragment antibodies show strong specificity in certain detection methods (such as immunohistochemistry, immunofluorescence, etc.) and can be used for high-sensitivity detection.
Does the recombinant fragment antibody customization service support multiple expression systems?

Yes, the recombinant fragment antibody customization service usually supports multiple expression systems, and customers can choose the appropriate system for antibody expression according to actual needs. Common expression systems include E. coli system, yeast system, insect cell system and mammalian cell system (such as CHO cells, HEK293 cells, etc.). Different expression systems have their own advantages and disadvantages. The E. coli system is the most commonly used efficient and low-cost expression system, but it may not be able to complete complex glycosylation modifications and is suitable for expressing simple antibody fragments. The yeast system can provide certain glycosylation modifications, but the expression level is not as high as that of E. coli. Insect cell systems are often used to express antibodies that require a certain degree of glycosylation, especially when polysaccharide modification requirements are high. Mammalian cell systems can provide complex glycosylation modifications and are suitable for the production of high-quality recombinant antibody fragments. Customers can choose the appropriate expression system based on the needs and application scenarios of antibodies.
What does quality control include in the customized service of recombinant fragment antibodies?

The quality control link in the customized service of recombinant fragment antibodies is very important, mainly including the following aspects: First, affinity and specificity testing is the core of quality control. Through ELISA, surface plasmon resonance (SPR), immunoblotting (Western Blot) and other methods, the binding ability of antibody fragments to target antigens is tested to ensure that they have high affinity and specificity. Secondly, purity testing is the key to ensuring the quality of antibodies. The purity of antibodies is tested by SDS-PAGE, HPLC and other technologies to ensure that there are no impurities and unnecessary proteins in the sample. Third, stability testing is also part of quality control, and it is necessary to evaluate the stability of antibodies under different conditions, including storage stability, freeze-thaw stability, etc. Fourth, immunogenicity evaluation is crucial for the clinical application of antibodies, especially in therapeutic applications, it is necessary to ensure that the immunoreactivity of antibody fragments is low. Finally, activity testing is also very important. The functional activity of the antibody is verified through cell experiments or animal experiments to ensure its effectiveness in practical applications.

What are recombinant fragment antibodies and why should we choose this form of antibody?

Recombinant fragment antibodies refer to recombinant antibodies composed of antibody fragments (such as Fab, scFv, Fv, etc.). Compared with traditional complete antibodies (IgG), recombinant fragment antibodies remove the constant region of the antibody and retain only the variable region, thus having a smaller molecular weight. This gives recombinant fragment antibodies some unique advantages, such as better tissue permeability, lower immunogenicity, and faster molecular clearance rate. In targeted therapy and diagnostic applications, recombinant fragment antibodies can more effectively enter tumor cells or other target tissues, so they are widely used in tumor immunotherapy, vaccine development, pathological detection, etc. Fragment antibodies produced by recombinant technology can achieve higher specificity and affinity, thus showing higher advantages in research and clinical applications.
What are the main differences between recombinant fragment antibodies and full-length antibodies?

The main difference between recombinant fragment antibodies and full-length antibodies lies in their structure and function. Full-length antibodies are usually composed of two heavy chains and two light chains, including variable regions and constant regions. The constant region plays an important role in the immune function of antibodies (such as ADCC, CDC, etc.), while fragment antibodies only retain the variable region of antibodies, that is, the antigen-binding fragment. This makes fragment antibodies small in size and simple in structure, and usually have no immune effector function, such as no activation of antibody-dependent cell-mediated cytotoxicity (ADCC) and classical complement-dependent cytotoxicity (CDC). The advantage of fragment antibodies is that they can better penetrate into tissues or cells, have faster serum clearance, and are generally less immunogenic than full-length antibodies. In addition, fragment antibodies are suitable for precision medicine and targeted therapy, such as cancer targeted drugs and molecular probes, while full-length antibodies are more used in treatments that require immune effects, such as the treatment of infections and autoimmune diseases.
What are the advantages of recombinant fragment antibody customization services?

Recombinant fragment antibody customization services have multiple advantages over traditional antibody preparation methods. First, fragment antibodies are smaller than full-length antibodies, have lower molecular weight, and have better tissue permeability, which is very important for applications such as tumor targeted therapy that require rapid penetration into cells or tissues. Secondly, fragment antibodies have reduced immunogenicity due to the removal of constant regions, so they may lead to fewer immune responses in clinical applications. In addition, the production of fragment antibodies is usually simpler and more efficient than full-length antibodies, which can reduce costs and time. Through genetic engineering technology, fragment antibodies can be optimized to improve their affinity, specificity and other properties, so as to customize antibodies that better meet the needs of customers. Finally, recombinant fragment antibodies show strong specificity in certain detection methods (such as immunohistochemistry, immunofluorescence, etc.) and can be used for high-sensitivity detection.
Does the recombinant fragment antibody customization service support multiple expression systems?

Yes, the recombinant fragment antibody customization service usually supports multiple expression systems, and customers can choose the appropriate system for antibody expression according to actual needs. Common expression systems include E. coli system, yeast system, insect cell system and mammalian cell system (such as CHO cells, HEK293 cells, etc.). Different expression systems have their own advantages and disadvantages. The E. coli system is the most commonly used efficient and low-cost expression system, but it may not be able to complete complex glycosylation modifications and is suitable for expressing simple antibody fragments. The yeast system can provide certain glycosylation modifications, but the expression level is not as high as that of E. coli. Insect cell systems are often used to express antibodies that require a certain degree of glycosylation, especially when polysaccharide modification requirements are high. Mammalian cell systems can provide complex glycosylation modifications and are suitable for the production of high-quality recombinant antibody fragments. Customers can choose the appropriate expression system based on the needs and application scenarios of antibodies.
What does quality control include in the customized service of recombinant fragment antibodies?

The quality control link in the customized service of recombinant fragment antibodies is very important, mainly including the following aspects: First, affinity and specificity testing is the core of quality control. Through ELISA, surface plasmon resonance (SPR), immunoblotting (Western Blot) and other methods, the binding ability of antibody fragments to target antigens is tested to ensure that they have high affinity and specificity. Secondly, purity testing is the key to ensuring the quality of antibodies. The purity of antibodies is tested by SDS-PAGE, HPLC and other technologies to ensure that there are no impurities and unnecessary proteins in the sample. Third, stability testing is also part of quality control, and it is necessary to evaluate the stability of antibodies under different conditions, including storage stability, freeze-thaw stability, etc. Fourth, immunogenicity evaluation is crucial for the clinical application of antibodies, especially in therapeutic applications, it is necessary to ensure that the immunoreactivity of antibody fragments is low. Finally, activity testing is also very important. The functional activity of the antibody is verified through cell experiments or animal experiments to ensure its effectiveness in practical applications.

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Recombinant Fragment Antibody Customization Service Frequently Asked Questions

What are recombinant fragment antibodies and why should we choose this form of antibody?

What are the main differences between recombinant fragment antibodies and full-length antibodies?

What are the advantages of recombinant fragment antibody customization services?