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- W1542392353 abstract "Publisher Summary Identification of the affinity ligand attachment sites on the B-chain of proteins like blocked ricin is a matter of interest in recent studies. In general, the identification of reactive or chemically modified residues of proteins is extremely important for the characterization of proteins and their activity. Ricin is a heterodimeric protein composed of a toxic A-chain, which is responsible for inhibiting cellular protein synthesis, disulfide-linked to a B-chain, known to possess lectin activity. Identification of the residues of ricin involved in the covalent linkage to the affinity ligand is important for complete characterization of the cytotoxic effector moiety, and to ensure consistency of the immunoconjugate product. However, because of the intrinsic heterogeneity of the ligand, isolation of individual species of ligandbound B-chain peptides by traditional peptide mapping has not been possible. To minimize the effect of the ligand heterogeneity, a different approach was conceived exploiting the incorporated protected thiol at the peptide “end” of the ligand. The thiol-specific probe, 2-(4 ′ -maleimidylanilino)naphthalene-6-sulfonic acid (MIANS) was used to label the ligand linked to B-chain in situ . MIANS is an attractive choice because its characteristic absorbance profile facilitates the identification of the peptides of interest. A monoclonal antibody recognizing MIANS is produced, such that affinity chromatography can be used to isolate only those peptides of the B-chain that are cross-linked to the MIANS-labeled ligand. This chapter discusses the results obtained using MIANS-labeling in conjunction with affinity chromatography to map free thiols in reduced, native ricin A-chain. It further describes the results obtained when utilizing this method to isolate ligand-bound ricin B-chain peptides." @default.
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- W1542392353 date "1997-01-01" @default.
- W1542392353 modified "2023-09-23" @default.
- W1542392353 title "The identification and isolation of reactive thiols in ricin A-chain and blocked ricin using 2-(4′-maleimidylanilino)naphathalene-6-sulfonic acid" @default.
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- W1542392353 doi "https://doi.org/10.1016/s1080-8914(97)80026-9" @default.
- W1542392353 hasPublicationYear "1997" @default.
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