کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1176935 | 961905 | 2011 | 7 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Quantitation of a recombinant monoclonal antibody in monkey serum by liquid chromatography–mass spectrometry Quantitation of a recombinant monoclonal antibody in monkey serum by liquid chromatography–mass spectrometry](/preview/png/1176935.png)
A method including protein A purification, limited Lys-C digestion, and mass spectrometry analysis was used in the study to quantify a recombinant monoclonal antibody in cynomolgus monkey serum. The same antibody that was isotopically labeled was used as an internal standard. Interferences from serum proteins were first significantly reduced by protein A purification and then by limited Lys-C digestion of protein A bound IgG, including both monkey and the recombinant IgG. Fab fragment of the recombinant human IgG was analyzed directly by LC–MS, while monkey IgG and the Fc fragment of the recombinant human IgG remained bound to protein A resin. Quantitation was achieved by measuring the peak intensity of the Fab from the recombinant human IgG and comparing it to that of the Fab from the stable isotope-labeled internal standard. The results were in good agreement with the values from ELISA. LC–MS can therefore be used as a complementary approach to ELISA to quantify recombinant monoclonal antibodies in serum for pharmacokinetics studies and it can also be used where specific reagents such as antigens are not readily available for ELISA.
Journal: Analytical Biochemistry - Volume 414, Issue 1, 1 July 2011, Pages 147–153