{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T04:06:36Z","timestamp":1791173196932,"version":"4.1.0"},"reference-count":30,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2004,1,29]],"date-time":"2004-01-29T00:00:00Z","timestamp":1075334400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2004,3,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>During production of a humanized antibody fragment secreted into the periplasm of <jats:italic>Escherichia coli<\/jats:italic>, proteolytic degradation of the light chain was observed. In order to determine which protease(s) were responsible for this degradation, we compared expression of the F(ab\u2032)<jats:sub>2<\/jats:sub> antibody fragment in several <jats:italic>E. coli<\/jats:italic> strains carrying mutations in genes encoding periplasmic proteases. Analysis of strains cultured in high cell density fermentations showed that the combination of mutations in <jats:italic>degP prc spr<\/jats:italic> was necessary for the cells to produce high levels of the desired recombinant antibody fragment. In order to eliminate the possible effects of mutations in other genes, we constructed <jats:italic>E. coli<\/jats:italic> strains with protease mutations in isogenic backgrounds and repeated the studies in high cell density fermentations. Extensive light chain proteolysis persisted in <jats:italic>degP<\/jats:italic> strains. However, light chain proteolysis was substantially decreased in <jats:italic>prc<\/jats:italic> and <jats:italic>prc spr<\/jats:italic> strains, and was further decreased with the introduction of a <jats:italic>degP<\/jats:italic> mutation in <jats:italic>prc<\/jats:italic> and <jats:italic>prc spr<\/jats:italic> mutant strains. These results show that the periplasmic protease Prc (Tsp) is primarily responsible for proteolytic degradation of the light chain during expression of a recombinant antibody fragment in <jats:italic>E. coli<\/jats:italic>, and that DegP (HtrA) makes a minor contribution to this degradation as well. The results also show that <jats:italic>spr<\/jats:italic>, a suppressor of growth defects in <jats:italic>prc<\/jats:italic> strains, is required for a <jats:italic>prc<\/jats:italic> mutant to survive throughout high cell density fermentations. \u00a9 2004 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/bit.20014","type":"journal-article","created":{"date-parts":[[2004,1,30]],"date-time":"2004-01-30T04:00:27Z","timestamp":1075435227000},"page":"463-474","source":"Crossref","is-referenced-by-count":70,"title":["High\u2010level accumulation of a recombinant antibody fragment in the periplasm of <i>Escherichia coli<\/i> requires a triple\u2010mutant (<i>degP prc spr<\/i>) host strain"],"prefix":"10.1002","volume":"85","author":[{"given":"Christina","family":"Chen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brad","family":"Snedecor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie C.","family":"Nishihara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John C.","family":"Joly","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nancy","family":"McFarland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dana C.","family":"Andersen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John E.","family":"Battersby","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen 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