Rapid Screening and Effective Rabbit-Derived Fab Antibodies Production Based on Yeast Surface Display.

INTRODUCTION/

OBJECTIVE

Antibodies have broad applications in various fields, such as biology and medicine. The screening and preparation of highly specific and sensitive antibodies are essential research areas. Several techniques for the preparation of mouse-derived antibodies have been developed, but limited studies on rabbit-derived antibodies with a broader antibody profile and easier humanization are reported. An improved yeast surface display technique was used for rapid screening of rabbit-derived Fab antibodies.

METHODS

After RNA extraction from peripheral rabbit blood, a cDNA library was obtained by reverse transcription. After recombinant vector construction, the expressed sequence in the form of Fab antibody structure was fused to the N-terminal end of Aga2p in the vector; a bidirectional promoter was inserted and successfully expressed in brewer's yeast EBY100. In addition, sequences, such as leucine zipper and inulinase signal peptide (INU), were inserted into the recombinant vector to improve the expression and stability of Fab antibody further.

RESULTS

A biotin-labeled salbutamol marker was synthesized, and two rabbit-derived salbutamol- Fab antibodies were screened in three weeks using fluorescence-activated cell sorting (FACS).

CONCLUSION

After antigen-binding kinetic studies, the screened antibodies demonstrated good affinity and specificity.