Abstract Lipocalin, a family of diverse proteins that distribute widely in organisms, exhibits several advantages, compared with antibody. To construct a high-capacity and good-diversity ribosome display anticalin library for selecting small molecular anticalin with high affinity, bilin-binding protein gene(bbp) that is a member of lipocalin family from Pieris brassicae, was employed for the preparation of a random library. A DNA library, named bbp gene library, encoding amino acids of bbp possessing 16 randomly mutated residues, was generated using gene splicing by overlap extension PCR (SOE-PCR). Then, using the same method the elements (T and P fragments) that were necessary for efficient transcription and translation in vitro were introduced to bbp gene library to construct the ribosome display anticalin library. In conclusion, we successfully constructed a ribosome display anticalin library with a volume of 3.76×1023, high-capacity and good-diversity. This library could be served as an efficient selection system for antibodies of micro-molecular haptens.
