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These enzymes catalyze the essential cross\u2010linking step of the polymerization of cell wall peptidoglycan. The understanding of the mechanisms of action of \u03b2\u2010lactams and of resistance to these drugs requires the development of reliable methods to characterize their targets. Here, we describe an activity\u2010based purification method of \u03b2\u2010lactam targets based on click and release chemistry. We synthesized alkyne\u2010carbapenems with suitable properties with respect to the kinetics of acylation of a model target, the Ldt\n                    <jats:sub>fm<\/jats:sub>\n                    L,D\u2010transpeptidase, the stability of the resulting acylenzyme, and the reactivity of the alkyne for the cycloaddition of an azido probe containing a biotin moiety for affinity purification and a bioorthogonal cleavable linker. The probe provided access to the fluorescent target in a single click and release step.\n                  <\/jats:p>","DOI":"10.1002\/chem.202100653","type":"journal-article","created":{"date-parts":[[2021,4,1]],"date-time":"2021-04-01T05:07:36Z","timestamp":1617253656000},"page":"7687-7695","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Click and Release Chemistry for Activity\u2010Based Purification of \u03b2\u2010Lactam Targets"],"prefix":"10.1002","volume":"27","author":[{"given":"Saidbakhrom","family":"Saidjalolov","sequence":"first","affiliation":[{"name":"Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, UMR 8601 CNRS Universit\u00e9 de Paris  45, rue des saints-p\u00e8res Paris 75006 France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuelle","family":"Braud","sequence":"additional","affiliation":[{"name":"Laboratoire de Chimie et Biochimie Pharmacologiques et 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