DOI: 10.3390/vetsci13080788 ISSN: 2306-7381

A Method Based on Nanobodies Against AMH to Establish Immunochromatography for Evaluation of Follicular Development in Bactrian Camels

Li Wang, Yaqian Jin, Guoli Zhang, Xue Chen, Hongrui Ren, Lanjie Li, Jingjing Tian, Bayier Mengke, Peiming Li, Yongjia Ma, Caolemeng Qiqige, Wa Gao, Guiqin Liu, Ruiwen Fan

(1) Background: Anti-Müllerian hormone (AMH) is a key indicator for predicting ovarian reserve and fertility. Currently, B-mode ultrasound examination is the primary method for evaluating follicular development in Bactrian camels, although it has several limitations. This study aims to develop a colloidal gold strip based on AMH nanobody preparation, establishing an efficient, simple, and rapid detection method suitable for evaluating follicular development. (2) Methods: Two nanobodies specifically binding to AMH were screened and identified from an alpaca naive nanobody library using phage display technology. The nanobodies were expressed in the yeast expression vector (pMCO-AOXα) and purified by affinity chromatography on a nickel column. Based on the purified nanobodies, a colloidal gold immunochromatographic strip was constructed for detecting serum AMH levels. (3) Results: In this study, anti-AMH nanobodies (AMH-VHH-A1 and AMH-VHH-A10) were expressed and purified; these were successfully utilized to prepare colloidal gold immunochromatographic strips for detecting AMH in serum. The T-line intensity of the colloidal gold detection was positively correlated with the number of growing follicles obtained by B-mode ultrasound imaging. (4) Conclusions: The colloidal gold strips based on nanobodies against AMH established in this research provided an effective, simple, rapid, and on-site method for evaluating follicular development, thus supplying a reference for reproductive management in Bactrian camels.

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