Design and Synthesis of Haptens for 3-amino-5-morpholinomethyl (AMOZ)

Creative Biolabs is a leading service provider that focuses on hapten discovery services. Currently, various platforms have been established for designing haptens in antibody production. Currently, we have successfully established a series of novel haptens for 3-amino-5-morpholinomethyl (AMOZ) to help develop small-molecule antibodies for our worldwide clients.

Introduction to 3-amino-5-morpholinomethyl (AMOZ)

AMOZ, the main type of furaltadone metabolite, has been considered as an attractive marker for evaluating the corresponding drug in disease therapy. Normally, AMOZ can be stably expressed and detected in patients, and thereby suggesting its essential role in monitoring the illegal use of furaltadone in edible tissues. Meanwhile, a number of methods, such as liquid chromatography with a mass spectrometer (LC-MS) and ultraviolet (UV), have been generated to detect AMOZ in different tissues. The results have shown that these assays can provide a rapid, accurate and sensitive screening process for quantifying the AMOZ. For example, many reports have revealed the broad use of enzyme-linked immunosorbent assay (ELISA) in the determination of furaltadone and AMOZ. Furthermore, recent studies have indicated that AMOZ can interact with o-nitrobenzaldehyde (o-NBA) to form the nitrophenyl (NP) derivative (NPAMOZ) prior to detection. As a result, the detection assay for AMOZ can also be designed by using antibodies targeting NPAMOZ.

Generally, the haptens for AMOZ can be produced based on carboxybenzaldehyle (CBA). These haptens can be coupled to different kinds of carrier proteins through a suitable length of the carboxylic acid spacer, which plays an important role in generating effective antibodies. For instance, an antibody R221 has been reported in a recent study, and the data have suggested that this antibody has a remarkable cross-reactivity (34.4%) with furaltadone and has a low cross-reactivity (2.3%) with AMOZ. The efficacy of R221 still needs to be improved by optimizing the structure of target molecules.

Synthesis of haptens against AMOZ. Fig.1 Synthesis of haptens against AMOZ. (Liu, 2018)

Our Platform for Design and Synthesis of AMOZ Haptens

Furaltadone has been regarded as a critical antibiotic for preventing and treating bacterial infections in livestock. However, due to the carcinogenic and mutagenic property of furaltadone, many attempts have been made to evaluate the potential risk in humans. In the past few years, a panel of assays has been developed for detecting its metabolite, AMOZ, in animal tissues. Moreover, a wide spectrum of immunoassay has been also generated for AMOZ based on antibodies specific to the derivative of AMOZ.

Currently, Creative Biolabs has developed a well mature and comprehensive hapten design and synthesis platform for the detection of AMOZ and the production of AMOZ-based antibodies. In our company, the immunizing haptens for AMOZ will be obtained by derivatizing AMOZ using specific acetic acid, 3-CBA, and 4-CBA. The potency of these novel haptens to produce effective antibodies against NPAMOZ will be assessed by using different assays, such as ELISA and LC-MS. The assay sensitivity will be further evaluated by using an optimal heterologous coating hapten. Moreover, the structures of NPAMOZ and the haptens can be also analyzed by molecular modeling to select the lowest energy conformation of target molecules and new haptens. Additionally, a battery of animals, including rats, mice, and rabbits, can be immunized with specific immunogens in our labs to obtain antibodies against NPAMOZ. Meanwhile, we also provide a number of services for analyzing the specificity and cross-reactivity of candidate antibodies.

Standard curves and calibration curves in the linear range (inset) of two antiserums for AMOZ. Fig.2 Standard curves and calibration curves in the linear range (inset) of two antiserums for AMOZ. (Liu, 2018)

As a reliable small-molecule antibody development company, Creative Biolabs is committed and proud to be part of this rewarding challenge by offering its support to the hapten discovery services and pre-clinical drug discovery for our clients. We are proud to partner with you on the journey of bringing novel haptens to disease therapy in the clinic. If you are interested in our services, please contact us for more details.

Reference

  1. Liu, F.; et al. Hapten synthesis and production of specific antibody against 3-amino-5-morpholinomethyl-2-oxazolidone for immunoassay without derivatization. Food and agricultural immunology. 2018, 29(1): 332-345.
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