Vance, Nicholas; Zacharias, Neelie; Ultsch, Mark; Li, Guangmin; Fourie, Aimee; Liu, Peter; LaFrance-Vanasse, Julien; Ernst, James A.; Sandoval, Wendy; Kozak, Katherine R.; Phillips, Gail; Wang, Weiru; Sadowsky, Jack published an article on January 16 ,2019. The article was titled 《Development, optimization, and structural characterization of an efficient peptide-based photoaffinity cross-linking reaction for generation of homogeneous conjugates from wild-type antibodies》, and you may find the article in Bioconjugate Chemistry.HPLC of Formula: 76931-93-6 The information in the text is summarized as follows:
Site-specific conjugation of small mols. to antibodies represents an attractive goal for the development of more homogeneous targeted therapies and diagnostics. Most site-specific conjugation strategies require modification or removal of antibody glycans or interchain disulfide bonds or engineering of an antibody mutant that bears a reactive handle. While such methods are effective, they complicate the process of preparing antibody conjugates and can neg. impact biol. activity. Herein we report the development and detailed characterization of a robust photoaffinity crosslinking method for site-specific conjugation to fully glycosylated wild-type antibodies. The method employs a benzoylphenylalanine (Bpa) mutant of a previously described 13-residue peptide derived from phage display to bind tightly to the Fc domain; upon UV irradiation, the Bpa residue forms a diradical that reacts with the bound antibody. After the initial discovery of an effective Bpa mutant peptide and optimization of the reaction conditions to enable efficient conjugation without concomitant UV-induced photodamage of the antibody, we assessed the scope of the photoconjugation reaction across different human and nonhuman antibodies and antibody mutants. Next, the specific site of conjugation on a human antibody was characterized in detail by mass spectrometry experiments and at at. resolution by X-ray crystallog. Finally, we adapted the photoconjugation method to attach a cytotoxic payload site-specifically to a wild-type antibody and showed that the resulting conjugate is both stable in plasma and as potent as a conventional antibody-drug conjugate in cells, portending well for future biol. applications.2,5-Dioxopyrrolidin-1-yl 2-(acetylthio)acetate(cas: 76931-93-6HPLC of Formula: 76931-93-6) was used in this study.
2,5-Dioxopyrrolidin-1-yl 2-(acetylthio)acetate(cas: 76931-93-6) belongs to pyrrolidine. Pyrrolidine being a good nucleophile easily undergoes electrophilic substitution reactions with different electrophiles such alkyl halides and acyl halides, and forms N-substituted pyrrolidines. N-Alkylpyrrolidine on further reaction with alkyl halide provided quaternary salts.HPLC of Formula: 76931-93-6
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