Description_x000D_ Application_x000D_ Multiplex Universal Peptide Quant Approach Using Microflow LC/MS/MS for Non-human Preclinical PK Study of Therapeutic Monoclonal Antibodies ( Webinar )_x000D_ Applying Best-in-Class HPLC Column Technologies to the Analysis of Proteins and Biotherapeutics Using SILuMab ( Webinar )_x000D_ Features and Benefits_x000D_ Universal Peptide Sequence Location_x000D_ YGPPCPPCPAPEFLGGPSVFLFPPKPK Heavy Chain (IgG4 Stabilized)_x000D_ VVSVLTVLHQDWLNGK Heavy Chain (IgG1, IgG3, IgG4)_x000D_ GFYPSDIAVEWESNGQPENNYK Heavy Chain (IgG1, IgG4)_x000D_ TTPPVLDSDGSFFLYSR Heavy Chain (IgG4)_x000D_ SGTASVVCLLNNFYPR Light Chain (kappa)_x000D_ VDNALQSGNSQESVTEQDSK Light Chain (kappa)_x000D_ DSTYSLSSTLTLSK Light Chain (kappa)_x000D_ Underlined is a Serine-Proline substitution in the hinge region that is present in many human IgG4-based therapeutic monoclonal antibodies_x000D_ SILuMab has been validated as an internal standard for quantitation of relevant biotherapeutics in a complex biological matrix by MRM-based LC-MS/MS._x000D_ • SILuMab yielded reproducible, linear curves from 0.1 μg/mL to 1000 μg/mL without enrichment or depletion._x000D_ • Good agreement was observed between multiple peptides derived from the same target._x000D_ • Label incorporation was determined to be >98% by mass spectrometry._x000D_ • Sequence coverage was confirmed by peptide mapping._x000D_ Physical form_x000D_ Supplied as a lyophilized powder containing phosphate buffered saline_x000D_ Preparation Note_x000D_ Produced utilizing enriched media containing stable isotope labeled amino acids are 13C6, 15N4-labeled arginine and 13C6, 15N2-labeled lysine_x000D_ SILuMab design is optimized to be used as an internal standard for quantitation of monoclonal antibodies as well as Fc-fusion therapeutics. Because of overlap with the common sequences in the Fc region with candidate antibodies, SILuMab provides univer­sal utility, thus eliminating the need for production of candidate-specific internal standards._x000D_ Reconstitution_x000D_ SILuMab recovery is maximized when 0.1% formic acid is used for reconstitution of the lyophilized product. Reconstitution with other solvents may reduce recovery. Do not freeze after reconstitution._x000D_ Procedure_x000D_ • Briefly centrifuge the vial at ~10,000 x g to collect the product at the bottom of the vial._x000D_ • Add 500 μL of purified water containing 0.1% formic acid to the vial._x000D_ • Mix the contents by gently inverting the vial a minimum of 5 times._x000D_ • Allow the vial to stand at room temperature for a minimum of 15 minutes and repeat mixing by inversion._x000D_ Analysis Note_x000D_ Quantitative_x000D_ MRM settings provided (xls)_x000D_ SILuMAb K4 Heavy Chain_x000D_ EVQLVESGGGLVQPGGSLRLSCVASGFTLNNYDMHWVRQGIGKGLEWVSKIGTAGDRYYAGSVKGRFTISRENAKDSLYLQMNSLRVGDAAVYYCARGAGRWAPLGAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG_x000D_ SILuMAb K4 Light Chain_x000D_ QSALTQPRSVSGSPGQSVTISCTGTSSDIGGYNFVSWYQQHPGKAPKLMIYDATKRPSGVPDRFSGSKSGNTASLTISGLQAEDEADYYCCSYAGDYTPGVVFGGGTKLTVLTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC_x000D_ Target overlap areas are underlined_x000D_ Package size based on protein content determined by A280 using an extinction coefficient (E0.1%) of 1.4_x000D_ Legal Information_x000D_ This product is licensed under U.S. Patent No. 7,396,688 and foreign counterparts from E. I. du Pont de Nemours and Company. The purchase of this product conveys to the buyer the nontransferable right to use the purchased amount of the product for research and development only, including services for a third party for consideration. The buyer cannot sell or otherwise transfer this product, its components or materials made using this product or its components to a third party. Information about licenses for excluded uses is available from: E. I. du Pont de Nemours and Company; Attn: Associate Director, Commercial Development; DuPont Experimental Station E268; 200 Powdermill Rd.; Wilmington, DE 19803; 1-877-881-9787 (voice), 1-302-695-1437 (fax), licensing@dupont._x000D_ SILu is a trademark of Sigma-Aldrich Co. LLC