Full
Outline
Veterinary drug residues, pesticides, mycotoxins and their metabolites are widely present in food and environmental matrices at trace levels and pose potential carcinogenic, teratogenic and endocrine-disrupting risks,thereby necessitating the development of sensitive, rapid and high-throughput screening methods. Immunoassays employing antibodies as recognition elements are widely used in food safety monitoring owing to their low cost,rapid response and suitability for on-site analysis; however, their analytical performance largely depends on the availability of monoclonal antibodies with high affinity and specificity. Owing to the dual immune mechanisms of high-frequency somatic hypermutation and gene conversion, rabbit antibody repertoires exhibit remarkable diversity and superior epitope resolution, making rabbits an important source of monoclonal antibodies against small-molecule food contaminants. At present, high-affinity antibodies used in food safety immunoassays are predominantly rabbit polyclonal antibodies, which suffer from inherent limitations such as batch-to-batch variability, complex epitope composition, and difficulties in standardization and long-term stable supply, thereby restricting the engineering and industrialization of immunoassay methods. In contrast, rabbit monoclonal antibodies offer distinct advantages,including high affinity, single-epitope specificity, excellent reproducibility and amenability to genetic and structural engineering, highlighting their significant potential in small-molecule immunorecognition. This review focused on the design of small-molecule haptens and rabbit immunization strategies. It reviewed the recent advances in rabbit monoclonal antibody discovery platforms, including hybridoma technology, display libraries and single B-cell–based approaches, for the generation of rabbit monoclonal antibodies against small molecules. This review compared the strengths and limitations of these platforms in terms of screening throughput, preservation of native heavy- and light-chain variable region pairing, control of false-positive clones and antibody engineering potential. It also summarized the applications of rabbit monoclonal antibodies against small molecules in enzyme-linked immunoassays, lateral flow immunoassays and electrochemical and optical immunosensors discussed the opportunities and challenges associated with complex matrix tolerance, multi-residue detection and methodological standardization. Finally, this review highlighted the continued development of single-cell omics, microfluidics and computer-aided antibody design, which was expected to facilitate the development of rabbit-derived high-affinity antibodies against small molecules through more standardized and scalable workflows, thereby providing more reliable immunorecognition elements for food safety and environmental monitoring.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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