Antibody Enzymatic Biotinylation

After biotin or avidin is combined with antibody molecules or markers, it will not affect the affinity of the former and will not change the characteristics of the antibody. Not all of the four active sites of avidin bind to the biotin residues attached to the antibody molecule, and the remaining free sites can still serve as receptors for another biotin-labeled protein. These characteristics are the basis and principle of BAS immunolabeling technology. At present, there are many biotinylation reagents that can be used for antibodies, and the research on enzymes that can promote antibody biotinylation is also more and more extensive. Currently, researchers employ a microbial enzyme, transglutaminase (MTGase), to facilitate biotinylation of antibodies. The biotinylation of antibodies will gradually become more precise, and a precise covalent conjugate will be formed, which will not be easily affected by temperature, pH, organic solvents and denaturants.

The Services

Alfa Chemistry researchers have long been concerned with the research on biotin modification and labeling to achieve biotinylation at precise sites. The antibody biotinylation services we can provide include but are not limited to:

• Antibody biotinylation at specific sites
• Enzymatic biotinylation assay

About enzymatic biotinylation

The biotin-avidin system is a typical detection method as follows:

• Biotin-avidin method (BA) (labeled avidin and biotin method (LAB)): biotinylate the specific antibody, the enzyme molecule can be labeled on the avidin molecule, the biotinylated antibody and the detected After the antibody is combined, the enzyme molecule is linked to the antigen molecule with the high affinity of BAS, and the antigen can be detected through the enzymatic reaction.
• The BAB method is also called the BRAB method. The specific antibody is biotinylated, the enzyme molecule is marked on the biotin, and then the free avidin bridge is used to combine the tested antigen, biotinylated antibody and enzyme-labeled biotin.
• The principle of the ABC method is basically the same as the BAB method, except that avidin and enzyme-labeled biotin need to form an ABC complex in a certain proportion. This network structure combines a large number of enzyme molecules. When avidin has not been saturated by enzyme-labeled biotin, the biotinylated antibody can bind to it.

What can we provide?

• Bio-Avidin system for efficient detection
• Professional and reliable analysis

Notes on sample handling:

• When there are influencing factors in the reaction mixture solution, it will affect the detection process
• Avoid inactivation during antibody biotinylation
• The surface of the antibody has a large number of amino groups, resulting in large steric hindrance, which will affect the biotinylation efficiency of the antibody
• There is a competitive relationship. During the modification or labeling process, biotin may also bind to other groups, such as carbonyl, thiol and phenol groups.
• After labeling and modification, the sample needs to be fully dialyzed, otherwise the participating biological groups will compete for the binding of biotinylated antibodies and avidin.

Alfa Chemistry researchers have mastered advanced technology and can carry out efficient biotinylated antibody labeling, so as to achieve accurate detection. If you are interested in our services, please contact us immediately.

Reference:

1. Andre. J.; et al. Use of microbial transglutaminase for the enzymatic biotinylation of antibodies. Journal of Immunological Methods. 2000, 240(1-2): 47-54.

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