SILu™MAB Stable-Isotope Labeled Universal Monoclonal Antibody Standard human
SIGMA/MSQC3 - recombinant, expressed in CHO cells
Synonym: Mass spectrometry standard, IgG; Mass spectrometry standard, Immunoglobulin-G; Stable isotope labelled IgG
Product Type: Chemical
UV trace and deconvoluted spectra resulting from intact mass analysis of the SILUMab standard. Calculations were based on the assumption that 99% label incorporation was achieved. Excellent agreement was seen between the calculated and observed molecular weight values.
Incorporation of stable labeled isotopes for representative SILUMab peptides. No unlabeled peptides were detectable; therefore, incorporation was considered to be >98%.
Calibration curves of three SILUMab peptides obtained by spiking 200 μg/mL into beagle plasma containing Erbitux at 6.4–250 μg/mL.
Extracted ion chromatogram (XIC) of representative peptides from the digested SILUMab. Data was obtained from a 250 fmol injection of a SILUMab tryptic digest. Using optimized overlap with common sequences in the Fc region of candidate antibodies, SILUMab provides universal utility, thus eliminating the need for production of candidate-specific internal standards.
| assay | ≥90% (SDS-PAGE) |
| packaging | vial of 100 μg (± 10% Lot-specific vial content given on certificate of analysis) |
| Quality Level | 200  |
| recombinant | expressed in CHO cells |
| shipped in | wet ice |
| storage temp. | −20°C |
| Analysis Note: | Qualitative Intact heavy and light chains (FASTA file )Quantitative MRM settings provided (Skyline , xls ) |
| Analysis Note: | SILuMab Heavy Chain EVQLVESGGGLVQPGGSLRLSCVAS GTAGDRYYAGSVKGRFTISRENAKD APLGAFDIWGQGTMVTVSS|ASTKG PEPVTVSWNSGALTSGVHTFPAVLQ HKPSNTKVDKKVEPKSCDKTHTCPP TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ RDELTKNQVSLTCLVKGFYPSDIAVEWESNG YSKLTVDKSRWQQGNVFSCSVMHEA SILuMab Light Chain QSALTQPRSVSGSPGQSVTISCTGT DATKRPSGVPDRFSGSKSGNTASLT VFGGGTKLTVL|GQPKAAPSVTLFP AWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS Target overlap areas are underlined |
| Analysis Note: | Package size based on protein content determined by A280 using an extinction coefficient (E0.1%) of 1.4 |
| Application: | SILu™MAB Stable-Isotope Labeled Universal Monoclonal Antibody Standard human has been used in a comparative study of automated antibody de novo sequencing. |
| Features and Benefits: | Universal Peptide Sequence Location DTLMISR Heavy Chain (IgG1, IgG2, IgG3, IgG4) FNWYVDGVEVHNAK Heavy Chain (IgG1) VVSVLTVLHQDWLNGK Heavy Chain (IgG1, IgG3, IgG4) NQVSLTCLVK Heavy Chain (IgG1, IgG2, IgG3, IgG4) GFYPSDIAVEWESNGQPENNYK Heavy Chain (IgG1, IgG4) AGVETTTPSK Light Chain (lambda) YAASSYLSLTPEQWK Light Chain (lambda) |
| Features and Benefits: | SILuMab has been validated as an internal standard for quantitation of relevant biotherapeutics in a complex biological matrix by MRM-based LC-MS/MS. • SILuMab yielded reproducible, linear curves from 0.1 μg/mL to 1000 μg/mL without enrichment or depletion. • Good agreement was observed between multiple peptides derived from the same target. • Label incorporation was determined to be >98% by mass spectrometry. • Sequence coverage was confirmed by peptide mapping. |
| General description: | SILu™Mab is a recombinant, stable isotope-labeled, human monoclonal antibody which incorporates [13C6, 15N4]-Arginine and [13C6, 15N2]-Lysine. Expressed in CHO cells, SILuMab is designed to be used as a universal internal standard for bioanalysis of monoclonal antibodies. Because of overlap with common sequences in the Fc region with candidate antibodies, SILuMab provides universal utility and thus eliminates the need to produce candidate-specific internal standards. SILuMab is an IgG1 antibody with lambda light chain, but contains peptide sequences common to other IgG isotypes. |
| Legal Information: | SILu is a trademark of Sigma-Aldrich Co. LLC |
| Legal Information: | 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.com. |
| Physical form: | Supplied as a lyophilized powder containing phosphate buffered saline |
| Preparation Note: | Produced utilizing enriched media containing stable isotope labeled amino acids are 13C6, 15N4-labeled arginine and 13C6, 15N2-labeled lysine |
| Preparation Note: | 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. |
| Preparation Note: | 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. Procedure • Briefly centrifuge the vial at ~10,000 x g to collect the product at the bottom of the vial. • Add 500 μL of purified water containing 0.1% formic acid to the vial. • Mix the contents by gently inverting the vial a minimum of 5 times. • Allow the vial to stand at room temperature for a minimum of 15 minutes and repeat mixing by inversion. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Purity | ≥90% (SDS-PAGE) |
| Storage Temp. | −20°C |
| UNSPSC | 23201100 |