Design and Synthesis of Haptens for Monensin

Monensin is a polyether antibiotic isolated from Streptomyces cinnamomea with a molecular weight of 671 Da. Monensin can control the proportion of volatile fatty acids in the rumen, reduce protein degradation, reduce feed dry matter consumption and improve nutrient utilization, so it is widely used as a ruminant feed additive. However, because excessive amounts of monensin will remain and threaten the food safety of humans, accurate detection methods are very necessary, which requires the quality and output of its specific antibodies. Creative Biolabs provides monensin hapten design and synthesis services that can be used as immunogens and produce antibodies.

Fig.1 Structure of Monensin.

Introduction to Monensin

Monomectin is usually in the form of sodium salt when it is produced and used as a feed additive. There is no group directly coupled with the carrier protein, so it cannot be directly synthesized into immunogens. It must be modified and given a carboxyl group to combine with the protein to form immunogens. This protein binding process involves the introduction or activation of functional groups on the hapten molecule, and depending on its nature, it is coupled to the carrier protein while ensuring the yield of the conjugate.

Monensin Hapten Design and Synthesis Service

Generally, only large molecules, infectious substances or insoluble foreign bodies can elicit an immune response in the body and produce specific antibodies, but haptens are generally relatively small molecules that only trigger immunity when connected to a large carrier (such as a protein) reaction. This immune response allows us to obtain antibodies with good specificity, which can bind small-molecule hapten itself in vitro to complete various immunoassay tests. To provide customers with effective immunogens, Creative Biolabs provides the design and synthesis service of monensin hapten. Our service mainly includes the following two parts:

• Synthesis and purification of mono succinyl monensin.
• Preparation of the monensin-protein conjugate.

To covalently bind monensin to BSA and OVA proteins, we tested a variety of different coupling methods and reagents. The best solution is to continuously form hemisuccinate and mixed anhydride to react with the amino group of the protein to form an amide bond and 4-carbon spacer. The amino groups of the BSA and OVA carriers used to prepare the monensin conjugate were replaced by 35% and 50%, respectively, which represents the optimal range of hapten immune response.

In subsequent validation, we immunized rabbits and mice with a small dose of monensin-BSA to obtain antibodies and verify their specificity. Our results demonstrated that rabbit and mouse antiserums strongly identified monensin-protein conjugates in indirect and competitive ELISA and, to a lesser extent, in competitive ELISA, also identified free monensin.

If you want to inquire more about our monensin hapten design and synthesis services, please contact us.