纳米抗体是目前已知的最小的具有完整功能的抗体单体，近年来已经在科学研究、重大疾病诊断和治疗、小分子污染物检测等领域得到了广泛应用，具有极大的科研潜力和广阔的应用前景。优化纳米抗体，尤其是小分子污染物纳米抗体的制备技术，一直是该领域的难点和重点。为了解决传统研制方法低效、低成功率、低多样性等缺点，本项目结合高通量测序（NGS）技术依次以氟虫腈、三唑磷为靶标，进行纳米抗体研制新方法的摸索和验证，建立一套全新的无需人工挑选、扩增和鉴定的小分子污染物纳米抗体研制技术体系，最终用于噻虫嗪纳米抗体的研制，为这套方法用于其他小分子污染物纳米抗体的研发打下坚实的基础。主要研究内容包括：1）小分子污染物半抗原设计、合成及人工抗原的制备；2）噬菌体展示纳米抗体文库构建和人工生物淘选；3）新方法的摸索；4）新方法的验证；5）新方法的应用。这套方法的建立还将促进纳米抗体在其它学科领域的研究和应用。

Nanobodies are the smallest antibody domain with complete function. Recently, they have been widely used in scientific research, diagnosis and treatment of serious illnesses, and detection of small molecule contaminants, etc., with great research potential and broad application prospects. Improvement of the isolation technology of nanobodies, especially small-molecule contaminants, has always been difficult and key in this field. Followed by fipronil and triazophos as targets, this project developed and verified a generation method of nanobodies combined with high-throughput sequencing (NGS) technology. The shortcomings of traditional development methods such as low efficiency, low success rate, and low diversity are solved. This new set of small molecule pollutant nanobody development technology system does not require manual selection, amplification and identification, and is ultimately used in the development of thiamethoxam nanobody, which lays the foundation for the development of other small molecule pollutant nanobodies. The main research contents include 1) Design and synthesis of small molecule pollutant hapten and preparation of artificial antigen; 2) Construction of phage display Nanobody library and artificial biopanning; 3) Exploration of the new system; 4) Validation of the new system; 5) Application of the new system. The development of this method will also promote the research and application of nanobodies in other fields.