Biotin Single Chain Antibody Fragments

Based on the high affinity of the proteins avidin and streptavidin for the biotin molecule, the biotin-avidin/streptavidin system is used in a wide range of biotechnological and diagnostic applications. Biotin can be attached to the surface of proteins to form biotinylated proteins that can bind to avidin/streptavidin. It can be used for protein labeling, immobilization, detection and purification. Antibody energy and biotinylation of residues can improve the binding ability, and biotinylated antibodies are commonly used reagents in current molecular diagnostics. A common method for preparing biotinylated antibodies is to combine active biotin derivatives with certain chemical groups on the surface of proteins. It is also possible to site-specifically biotinylate the recombinant antibody, which can be linked to a specific group of a specific protein using a biotin ligase. Antibodies are composed of immunoglobulin domains, each of which contains an intradomain disulfide bond that plays an important role in stabilizing the protein structure. Functionalized single-strand biotinylated fragments can be prepared by a simple preparation method and can be used in high-efficiency and sensitive immunoassay applications.

The Services

Alfa Chemistry researchers are dedicated to the study of biotinylation at specific sites and can develop a variety of biotinylation methods that can be applied to a wide range of applications. The related services that our researchers can provide include but are not limited to:

• Construction of expression vector
• Optimization of biotin concentration
• Expression, purification and characterization of fusion proteins
• Determination of biotinylation concentration
• TSH immunoassay

About scFv

Single-chain antibody (scFv) is a small molecule composed of the variable region of the antibody heavy chain and the variable region of the light chain connected by a peptide chain, and is the smallest functional structural unit with antibody activity. Due to its small molecular weight, strong penetrating ability, and low immunogenicity, single-chain antibodies are widely used in the fields of clinical medicine, biological diagnosis, and treatment. On the basis of the complete antibody, the antibody heavy chain variable region (VH) and light chain variable region (VL) are connected into a recombinant gene by processing, so as to realize the expression of the recombinant gene. Fragments of single-chain antibodies are monovalent, and bivalent scFv can be produced by linking two scFv fragments. Bivalent scFv can form tandem antibodies or multimers with more antigen-binding sites. Single-chain antibodies can be expressed in multiple expression systems, the common ones are E. coli expression system and mammalian expression system.

Production of scFv

• cDNA extraction from hybridoma cells
• Distinguishing the heavy chain variable region VH of an antibody from the light chain variable region V_L
• Splicing of V_H and V_L fragments to obtain single-chain antibody variable region gene fragments (scFv)
• scFv cloned on phage to obtain single-chain antibody library
• Antigen-specific single-chain antibodies obtained by adsorption and elution

Figure 2. Preparation process and method of scFv

Wide application of scFv

• Wide application of scFv
• Targeted therapy
• Imaging diagnosis
• Intracellular immunity
• Biological detection

Alfa Chemistry has a complete set of antibody development solutions, including steps such as immunogen preparation, animal immunization, scFv antibody library construction and screening. Our researchers have many years of experience in the expression and purification of scFv antibodies, and can provide customers with high-quality services. If you are interested in our services, please contact us immediately.

Reference:

1. Santala. V.; et al. Production of a biotinylated single-chain antibody fragment in the cytoplasm of Escherichia coli. Journal of Immunological Methods. 2004, 284(1-2): 165-175.

※ It should be noted that our service is only used for research, not for clinical use.

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