Creative Biolabs has established a solid high-throughput sequencing analysis platform for Fab library sequence analysis. There are several approaches to identify a possible binder from a Fab library after the screening. One approach is to isolate and sequence the positive clones from the library by Sanger sequencing. Another approach is to use next-generation sequencing method for identification. However, both methods have their drawbacks. Creative Biolabs has developed an advanced Magic™ Fab repertoire analysis method, which could facilitate the antibody development in a much more efficient and effective manner.
What is Fab?
The Fab fragment is a region of an antibody that removes the Fc portion but still has the ability to bind to antigens. It consists of a constant and a variable domain that both have the heavy and light chain. The variable domain has the antigen-binding site with a set of complementarity-determining regions (CDRs) at N-terminus of the fragment. Fab fragment includes two antigen-binding portions which are linked by the disulfide bond. Therefore, each arm of Fab could bind to an epitope on the antigen. The fragment secondary antibody that recognizes the primary antibody is optimal for IHC applications.
Advantages of Magic™ Fab repertoire analysis
Magic™ Fab repertoire analysis is an advanced method to review the sequence information for the antigen-binding site of the antibody. It could dramatically improve the efficiency of screening and also be used to design new systems for novel antibody drugs discovery.
The traditional NGS method has a couple of drawbacks such as low quality of data, bias on detection of less frequent CDRs, and no information of the VH:VL pairing. Nevertheless, our Magic™ analysis system could overcome the limitations above. It introduces an engineering service which allows the sequencing of the paired antibody heavy and light chain variable regions. Furthermore, it minimizes the linker sequence, resulting in a deeper coverage of Fab libraries compared to the traditional Sanger sequencing. In addition, during the phage display selection, Magic™ analysis could verify the diversity of the initial library, which avoids the possibility of inherent propagation bias resulting from the multiple rounds of biopanning which is required by the traditional sequencing strategies.
Creative Biolabs is a world-class antibody development service provider. We offer one stop solution for Fab generation. From antigen synthesis, phage display library construction to screening and sequence analysis, we could provide tailored service to meet our customers’ unique requirement. With the Magic™ Fab repertoire analysis, Creative Biolabs has a proven record of generating Fab fragment with strong affinity and high selectivity in a timely manner.