Merck Supelco (Sigma-Aldrich) / BIOshell

BIOshell

BIOshell columns from Merck Supelco consist of core-shell particles for bioseparations ranging from glycans to large proteins. These columns are compatible with HPLC and UHPLC systems as well as for MS detection.
 

BIOshell IgG 1000 Å: for mAbs, ADCs, biosimilars, H/D exchange, mAb fragments

BIOshell A400 Protein: mAbs, ADCs, biosimolars, Proteins, mAb fragments

BIOshell A160 Peptide: for tryptic digest, post-translational modifications, polypeptides

BIOshell Glycan: for oligosaccharides via HILIC

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Technical Data

BIOshell IgG 1000 Å

Phase

Modification

Particle size

Pore size

USP

Tmax

BIOshell IgG 1000 Å C4

C4

2.7 μm

1000 Å

L26

90 °C

BIOshell IgG 1000 Å C18

C18

2.7 μm

1000 Å

L1

90 °C

BIOshell IgG 1000 Å Diphenyl

Diphenyl

2.7 μm

1000 Å

L11

90 °C

BIOshell A400 Protein

Phase

Modification

Particle size

Pore size

USP

Tmax

BIOshell A400 Protein C4

C4

3.4 μm

400 Å

L26

90 °C

BIOshell A400 Protein C18

C18

3.4 μm

400 Å

L1

90 °C

BIOshell A160 Peptide

Phase

Modification

Particle size

Pore size

USP

Tmax

pH range

BIOshell A160 Peptide Phenyl-Hexyl

Hexyl-Ph

2.7 μm

160 Å

L11

90 °C

1.0-8.0

BIOshell A160 Peptide C18

C18

2, 2.7, 5 μm

160 Å

L1

90 °C

1.0-8.0

BIOshell A160 Peptide Cyano

CN

2.7, 5 μm

160 Å

L10

90 °C

1.0-8.0

BIOshell Glycan

Phase

Modification

Particle size

Pore size

Carbon content

Surface area

USP

Tmax

BIOshell Glycan

proprietary pentahydroxy ligand

2.7 μm

90 Å

3.2 %C

135 m²/g

L95

65 °C

Applications

Effect of Phase Chemistry on Protein Selectivity

Measurement of denosumab with different columns.

 

Monoclonal antibodies are unique molecules and therefore can interact differently with different phase chemistries.
The numbered peaks correspond to IgG2 disulfide bond variants.

Test Conditions:

Columns: BIOshell™ IgG 1000 Å C4, 15 cm x 2.1 mm I.D., 2.7 μm

                  BIOshell™ IgG 1000 Å C18, 15 cm x 2.1 mm I.D., 2.7 μm

                  BIOshell™ IgG 1000 Å Diphenyl, 15 cm x 2.1 mm I.D., 2.7 μm

Mobile Phase A: 2:10:88 n-propanol/acetonitrile/water (0.1% v/v difluoroacetic acid

Mobile Phase B: 70:20:10 n-propanol/acetonitrile/water (0.1% v/v difluoroacetic acid)

Gradient: 16% B to 26% B in 20 min

Flow rate: 0.2 mL/min

Temperature: 80°C

Detection: UV, 280 nm

Injection volume: 2 µL

Sample: Denosumab, 2 mg/mL, water (0.1% v/v trifluoroacetic acid)

Rapid Protein Separations on BIOshell™ A400

Rapid Protein Separations on BIOshell™ A400.

 

Due to the shallow, porous shell, protein separations can be performed in less than one minute with the BIOshell™ A400 column.

1. lysozyme; chicken  14.3 kDa

2. haptoglobulin, phenotype 1-1; human 96 kDa

3. glutamate dehydrogenase; bovine  332 kDa

4. ß-galactosidase; E. coli   465 kDa

5. lactate dehydrogenase contaminant

6. lactate dehydrogenase; rabbit  140 kDa

Different Phase Chemistries Allows for Alternative Selectivity for Peptides

Measurement of amino acid sequences with different columns.

 

Better sequence coverage, through better resolution of critical pairs, can be achieved by using phase chemistries orthogonal to C18.

1. FTISADTSKNTAYLQMNSLR

2. LSCAASGFNIKDTYIHWVR

3. GFYPSDIAVEWESNGQPENNYK

4. LLIYSASFLYSGVPSR

5. SGTASVVCLLNNFYPR

6. SCDKTHTCPPCPAPELLGGPSVFLFPPKPK

7. VVSVLTVLHQDWLNGKEYK

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