The Research On The Applications Of Nanobody In Traditional Antibody Affinity Purification

Highly purified IgG antibody has been widely applicated in biological detection, clinical diagnosis and treatment of diseases. The common methods of the purification of IgG antibody include precipitation and affinity chromatography. Currently, the most widely used technique in laboratory and industry is the application of Protein A/G, which specifically binds to the Fc fragment of antibody. However, there are still some improvements in the affinity and specificity for Protein A/G binding to antibody. This research is based on the high affinity and specificity for antigen binding to antibody. Animals were immunized with Fc fragment of human antibody, phage display library with variable region of camel heavy chain antibody was constructed, and highly affinity and specific nanobody which binds to Fc fragment of human antibody were screened out through phage display technology. The nanobody was largely expressed, purified and linked to the agarose matrix, which can be used for the antibody affinity purification. The results demonstrate that compared with Protein A/G, anti-Fc nanobody has higher specificity which can bind to antibody with higher purification. In addition, the anti-Fc nanobody can also be applied for the fusion protein when doing affinity purification of Fc fragment, which provides a new way for this protein expression and purification.