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SILuLite SigmaMAb Universal Antibody Standard human

Synonym: IgG1 light, SILuLite SigmaMAb Universal Antibody Standard human, SigmaMAb

  •  NACRES NA.12

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Description

General description

SILU Lite SigmaMAb is a recombinant human monoclonal IgG1 lambda light antibody with a molecular mass of ∼150 kDa expressed in CHO cells. It is designed for optimization of accurate intact mass analysis of monoclonal antibodies, biosimilars, and pharmaceutical products. Accurate intact mass analysis of such large biomolecules can provide comprehensive information about structural and post-translational modifications such as glycosylation. Other information such as heterogeneity, batch-to-batch variation, amino acid truncation, and N-terminal Lys processing, aggregation, and degradation can be determined. Intact mass analysis using mass spectrometry is also very important for formulation and storage in therapeutic monoclonal antibody drug development.
It consists of two identical heavy chains and two identical light chains. The heavy chains and light chains are linked by one disulfide bond. The heavy chains are linked by two disulfide bonds located in a hinge domain. The other 12 cysteine bonds are intramolecularly restricted to six different globular domains. The antibody sequence has been evaluated by intact mass and peptide mapping using four different enzymes: chymotrypsin, Asp-N and Glu-C endoproteinases and trypsin. Sequence coverage of 100% was obtained.

Application

SILu Lite SigmaMAb Universal Antibody Standard human has been used as a model system to investigate the quantitative relationship between gas-phase monoclonal antibody (mAb) unfolding and the discrete levels of mAb glycosylation.

Features and Benefits

Calculated molecular weight values of the SigmaMAb light chains, heavy chains, and intact protein, with the most abundant glycoforms, are as follows:

Description / Composition / Modification / Average Mass (Da)

Light chain, reduced / C1006H1555N267O333S7 / Pyroglutamic acid (Q) / 22942.2

Heavy chain, reduced / C2181H3393N587O663S16 / (no modification) / 48957.8
C2237H3485N591O702S16 / G0F / 50403.2
C2243H3495N591O707S16 / G1F / 50565.3
C2249H3505N591O712S16 / G2F / 50727.5

Native intact mass, non-reduced / C6374H9864N1708O1992S46 / 2 X Pyroglutamic acid (Q) / 143767.7
C6486H10048N1716O2070S46 / G0F+G0F / 146658.4
C6492H10058N1716O2075S46 / G0F+G1F / 146820.6
C6498H10068N1716O2080S46 / G1F+G1F / 146982.7
C6504H10078N1716O2085S46 / G1F+G2F / 147144.8
C6510H10088N1716O2090S46 / G2F+G2F / 147307.0

Physical form

Supplied as a lyophilized powder containing phosphate buffered saline

Preparation Note

SigmaMAb recovery is maximized when phosphate buffer, pH 6–7, is used to reconstitute the lyophilized product.

Analysis Note

SigmaMab Heavy Chain
EVQLVESGGGLVQPGGSLRLSCVASGFTLNNYDMHWVRQGIGKGLEWVSKI
GTAGDRYYAGSVKGRFTISRENAKDSLYLQMNSLRVGDAAVYYCARGAGRW
APLGAFDIWGQGTMVTVSS|ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF
PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN
HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS
RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

SigmaMab Light Chain
QSALTQPRSVSGSPGQSVTISCTGTSSDIGGYNFVSWYQQHPGKAPKLMIY
DATKRPSGVPDRFSGSKSGNTASLTISGLQAEDEADYYCCSYAGDYTPGV
VFGGGTKLTVL|GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTV
AWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ
VTHEGSTVEKTVAPTECS

Also available as a stable isotope-labled product, SiluMab (Product Number MSQC3)

Package size based on protein content determined by A280

Other Notes

Avoid PBS for reconstitution.
Reconstitute the contents of the vial by adding 500μL of ultrapure water or phosphate buffer, and mixing vigorously. The solubilized product can be further diluted as needed.

Legal Information

SILu is a trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

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Certificate of Origin (COO)

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Protocols & Articles

Articles

A Rapid, Streamlined Workflow for Glycan Sample Preparation and Analysis by LC-MSpngase fast asms

Comparison of PNGase F workflows for glycan release. The traditional workflow (top) involves multiple wash steps and an overnight digestion. With PNGase Fast (bottom), digestion is complete in 15 min...
Judy Boland, Amber Henry, Nicolas Caffarelli, Gordon Nicol, Jeffrey L. Turner, and Kevin Ray
Keywords: Aminations, Detergents, Digestions, Electrophoresis, High performance liquid chromatography, Mass spectrometry, PAGE, Reductions, Reductive aminations

ADC Mimic Enables LC-MS Method Development

A non-toxic Cys-conjugated ADC-mimic characterized by top-down, middle-out, and bottom-up LC-MS methods.
Kevin Ray, Ben Cutak, Brian Gau
MilliporeSigma, St. Louis, MO
Keywords: Alkylations, Bacterial conjugations, Chromatography, Genetics, High performance liquid chromatography, Liquid chromatography mass spectrometry, Mass spectrometry, Reductions, Size-exclusion chromatography

ADC-Mimic for LC-UV-MS Method Development

Antibody-drug conjugates (ADC) represent a rapidly emerging class of biotherapeutic molecules, which harness the specificity of monoclonal antibodies (mAb) to selectively deliver highly potent cytoto...
Benjamin Cutak, Brian Gau, Nicolas Caffarelli, and Kevin Ray
MilliporeSigma, St. Louis, MO
Keywords: Bacterial conjugations, Chromatography, Fractionation, High performance liquid chromatography, Mass spectrometry, Reductions, Separation

BIOshell™ IgG 1000 Å C4 UHPLC Column for Improved Biomacromolecule Separations

Monoclonal antibodies (mAbs) are widely manufactured by many biopharmaceutical companies to treat a myriad of diseases ranging from Alzheimer’s disease, Parkinson’s disease, ulcerative colitis, and m...
Keywords: Amplification, Chromatography, Diffusion, Diseases, High performance liquid chromatography, Mass spectrometry, Methylations, Microscopy, Oxidations, Pesticides, Pharmaceutical, Phosphorylations, Reversed-phase chromatography, Scanning electron microscopy, Solid phase peptide synthesis

Fast UHPLC Analysis for IdeS Fragments of SiLu™Lite SigmaMAb Universal Antibody Standard on BIOshell A400 Protein C4

Both top-down and bottom-up approaches to protein characterization have their advantages and limitations. One such limitation of the top-down approach, particularly with large proteins, is the need f...
Hillel Brandes, and Stacy Shollenberger
Reporter US Volume 34.1
Keywords: Chromatography, Mass spectrometry, Reductions, Reversed-phase chromatography, Separation

Fast UHPLC Analysis of SILu™ Lite SigmaMAb Universal Antibody Standard IdeS Fragments on BIOshell™ A400 Protein C4

From our library of Articles, Sigma-Aldrich presents Fast UHPLC Analysis of SILu™ Lite SigmaMAb Universal Antibody Standard IdeS Fragments on BIOshell™ A400 Protein C4
Keywords: Chromatography, Immobilization, Mass spectrometry, Reductions, Reversed-phase chromatography, Sample preparations, Separation

HPLC Analysis of B Vitamins on Ascentis® Express RP-Amide, Acetonitrile Mobile Phase

From our library of Articles, Sigma-Aldrich presents HPLC Analysis of B Vitamins on Ascentis® Express RP-Amide, Acetonitrile Mobile Phase
Keywords: Chromatography, High performance liquid chromatography, Vitamins

HPLC Analysis of Oxidized SILu Lite SigmaMAb Universal Antibody Standard on BIOshell A400 Protein C4

From our library of Articles, Sigma-Aldrich presents HPLC Analysis of Oxidized SILu Lite SigmaMAb Universal Antibody Standard on BIOshell A400 Protein C4
Keywords: Chromatography, Oxidations, Reversed-phase chromatography, Separation

HPLC Analysis of Reduced SILu™ Lite SigmaMAb on Zenix® SEC-300 with pH-Neutral Buffer

From our library of Articles, Sigma-Aldrich presents HPLC Analysis of Reduced SILu™ Lite SigmaMAb on Zenix® SEC-300 with pH-Neutral Buffer
Keywords: Chromatography, High performance liquid chromatography, Pharmaceutical, Separation, Size-exclusion chromatography

HPLC Analysis of SILu Lite Antibody Standard on BIOshell™ A400 Protein C4 Versus Competitor Column

From our library of Articles, Sigma-Aldrich presents HPLC Analysis of SILu Lite Antibody Standard on BIOshell™ A400 Protein C4 Versus Competitor Column
Keywords: Chromatography, High performance liquid chromatography, Separation

HPLC Analysis of a Monoclonal Antibody Tryptic Digest on BIOshell™ A160 Peptide C18, 2.7 μm

From our library of Articles, Sigma-Aldrich presents HPLC Analysis of a Monoclonal Antibody Tryptic Digest on BIOshell™ A160 Peptide C18, 2.7 μm
Keywords: Alkylations, Chromatography, High performance liquid chromatography, Nucleic acid denaturation, Sample preparations

HPLC of monoclonal antibody (mAb) SigmaMab on BIOshell™ A400 Protein C4

From our library of Articles, Sigma-Aldrich presents HPLC of monoclonal antibody (mAb) SigmaMab on BIOshell™ A400 Protein C4
Keywords: High performance liquid chromatography, Separation

Non-Toxic ADC-Mimic for LC-MS Method Development

ADCs conjugated at Cys residues of inter-chain disulfide bonds give mAbs bearing 0-4 pairs of drug-linkers. Characterizing this distribution is paramount to understanding ADC efficacy and process con...
Brian Gau, Ben Cutak, Pegah Jalili, Kevin Ray
MilliporeSigma, St. Louis, MO
Keywords: Bacterial conjugations, Buffers, Filtration, High performance liquid chromatography, Liquid chromatography mass spectrometry, Mass spectrometry, Purification, Size-exclusion chromatography

Quantification of Host Cell Proteins in Biotherapeutics using Stable Isotope Labeled Chinese Hamster Ovary Proteins (SIL-CHOP) as an Internal Standard

Residual presence of host cell proteins (HCPs) in recombinant therapeutic products has considerable clinical safety risks associated with a potential immunological response in patients. Although trad...
Keywords: BCA assay, Cell culture, Clinical, Enzyme-linked immunosorbent assay, High performance liquid chromatography, Liquid chromatography mass spectrometry, Mass spectrometry, Purification, Solid phase extractions

UHPLC Analysis of Deglycosylated SigmaMAb Tryptic Digest on BIOshell A160 Peptide C18

From our library of Articles, Sigma-Aldrich presents UHPLC Analysis of Deglycosylated SigmaMAb Tryptic Digest on BIOshell A160 Peptide C18

UHPLC Analysis of SigmaMAb Universal Antibody Standard on BIOshell™ IgG 1000 Ã… C4

From our library of Articles, Sigma-Aldrich presents UHPLC Analysis of SigmaMAb Universal Antibody Standard on BIOshell™ IgG 1000 Ã… C4

Related Content

Stable Isotope-labeled MS Internal Standards

What happens when you combine state-of-the-art technology with scientific prowess? Full-length protein standards with the highest purity and isotopic incorporation in the industry. Stable isotope-lab...
Keywords: Atomic absorption spectroscopy, Buffers, Digestions, Mass spectrometry, PAGE, Phosphorylations

Peer-Reviewed Papers
15

References

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