{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T22:59:30Z","timestamp":1791413970815,"version":"4.3.3"},"reference-count":28,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2010,12,28]],"date-time":"2010-12-28T00:00:00Z","timestamp":1293494400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2011,1,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Recently, we have developed liquid chromatography\/tandem mass spectrometry (LC\/MS\/MS)\u2010based methods for the quantitation of pegylated therapeutic proteins in plasma. The methods are based on the LC\/MS\/MS detection of a surrogate peptide generated from trypsin digestion of the therapeutic protein. Various parameters related to the bioanalytical methods were evaluated and optimized, including the preparation of calibration standards and quality control samples, sample extraction, internal standard selection and its stage of addition, trypsin digestion, and non\u2010specific binding. In this paper, we report the development of a method for a specific pegylated therapeutic protein and detail the various optimization steps undertaken. Simple extraction of the pegylated therapeutic protein from plasma was achieved via the precipitation of the endogenous proteins in plasma using acidic isopropanol and the resulting supernatant extract was subjected to trypsin digestion. A unique tryptic peptide arising from the pegylated therapeutic protein was used for LC\/MS\/MS\u2010based detection and quantitation. A protein and a peptide were used as internal standards, with the former added before the sample extraction and the latter after the sample extraction. The method developed is simple, sensitive, specific and rugged, and has been implemented in a high throughput 96\u2010well format to analyze plasma samples from\n                    <jats:italic>in vivo<\/jats:italic>\n                    studies. A required lower limit of quantitation (LLOQ) of 10\u2009ng\/mL, expressed in terms of the concentration of the protein drug, was easily achieved. Copyright \u00a9 2010 John Wiley &amp; Sons, Ltd.\n                  <\/jats:p>","DOI":"10.1002\/rcm.4856","type":"journal-article","created":{"date-parts":[[2010,12,28]],"date-time":"2010-12-28T23:54:06Z","timestamp":1293580446000},"page":"281-290","source":"Crossref","is-referenced-by-count":68,"title":["A strategy for liquid chromatography\/tandem mass spectrometry based quantitation of pegylated protein drugs in plasma using plasma protein precipitation with water\u2010miscible organic solvents and subsequent trypsin digestion to generate surrogate peptides for detection"],"prefix":"10.1002","volume":"25","author":[{"given":"Steven T.","family":"Wu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Ouyang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy 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