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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 
 
 
 
 

Production and characterization of polyclonal anti-S 20499 antibodies: influence of the hapten structure on stereospecificity.

Immunoassays were studied as an alternative to HPLC methods for the stereoselective determination of a chiral drug, S 20499, a new anxiolytic compound that is chemically related to buspirone. The production of highly stereospecific polyclonal antibodies was sought following the construction of appropriately optimized hapten-protein conjugates. This process involved the selection of the structure and the length of the spacer arm used to couple S 20499 to the carrier protein as well as deciding on the location of the coupling site with respect to the chiral center. Two haptens were prepared: one a derivative resembling the original structure of S 20499, with the effective addition of a carboxylic acid group, and a second with the effective addition of a butanoic acid moiety that is supposed to favor stereorecognition. Six stereospecific polyclonal antisera were obtained in rabbits with two groups of antibody families defined in terms of specificity. Both approaches gave high levels of stereospecificity (cross-reactivity towards the optical antipode of S 20499 ranged from 4.1% to < 0.1%). Although it did not decrease the mean apparent affinity constant, the longer spacer improved antibody specificity by decreasing cross-reactions towards dealkylated S 20499 derivatives. Hence, the addition of a four carbon atom bridge should be a valuable tool for increasing antibody stereospecificity with no drawbacks in terms of specificity and affinity. It was also shown that long immunization periods appear to have no effect on the stereospecificity of the antibodies obtained.[1]

References

  1. Production and characterization of polyclonal anti-S 20499 antibodies: influence of the hapten structure on stereospecificity. Got, P.A., Guillaumet, G., Boursier-Neyret, C., Scherrmann, J.M. Journal of pharmaceutical sciences. (1997) [Pubmed]
 
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