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Proteins

Proteins are essential macromolecules that occur in all living organisms and fulfil a variety of biological functions, including enzyme activity, signal transmission and structure formation. Their precise analysis is crucial for the understanding of biochemical processes and the development of drugs and biotechnological products. High performance liquid chromatography (HPLC) is a key method for analysing proteins as it enables the precise separation and identification of different protein molecules. HPLC allows proteins to be broken down by size, charge or other chemical properties, making it an indispensable tool in biochemical research and the pharmaceutical industry.

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BioResolve SEC mAb Column with BioResolve SEC mAb Guard, 200Å, 2.5 µm, 7.8 x 300 mm, 1/pk with mAb Size Variant Standard;
176004599
€3,710.00
BioResolve SEC mAb Column with BioResolve SEC mAb Guard, 200Å, 2.5 µm, 7.8 x 150 mm, 1/pk with mAb Size Variant Standard;
176004598
€2,690.00
BioResolve SEC mAb Column with BioResolve SEC mAb Guard, 200Å, 2.5 µm, 4.6 x 300 mm, 1/pk with mAb Size Variant Standard;
176004597
€3,550.00
BioResolve SEC mAb Column with BioResolve SEC mAb Guard, 200Å, 2.5 µm, 4.6 x 150 mm, 1/pk with mAb Size Variant Standard;
176004596
€2,530.00
BioResolve SEC mAb Column, 200Å, 2.5 µm, 7.8 x 300 mm, 1/pk with mAb Size Variant Standard;
176004595
€3,610.00
BioResolve SEC mAb Column, 200Å, 2.5 µm, 7.8 x 150 mm, 1/pk with mAb Size Variant Standard;
176004594
€2,480.00
BioResolve SEC mAb Column, 200Å, 2.5 µm, 4.6 x 300 mm, 1/pk with mAb Size Variant Standard;
176004593
€3,430.00
BioResolve SEC mAb Column, 200Å, 2.5 µm, 4.6 x 150 mm, 1/pk with mAb Size Variant Standard;
176004592
€2,300.00
IEX-MS Start Up Kit: BioResolve SCX mAb 3 µm Column (2.1 x 100 mm) plus Humanized mAb Mass Check Standard plus IonHance CX-MS pH Buffer Concentrates in MS Certified LDPE Containers;
176004502
€2,040.00
IEX-MS Start Up Kit: BioResolve SCX mAb 3 µm Column (2.1 x 100 mm) with VanGuard FIT plus Humanized mAb Mass Check Standard plus IonHance CX-MS pH Buffer Concentrates in MS Certified LDPE Containers;
176004501
€2,130.00
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Technical Data

The chromatographic analysis of proteins requires different methods that are used depending on the physico-chemical properties of the proteins. Here are the most important chromatographic methods used to analyse proteins:

 

1. Reversed phase chromatography

  • Principle: In RP-HPLC, proteins are separated based on their hydrophobicity. Hydrophobic proteins bind more strongly to the column and elute later.
  • Application: This method is well suited for the separation of proteins and peptides that have different hydrophobicities. It is frequently used in protein purification and peptide sequencing.
  • Advantage: High resolution and good reproducibility.
  • Disadvantage: Denaturation of proteins by the organic solvents is possible.

 

2. Size exclusion chromatography (SEC)

  • Principle: SEC separates proteins based on their size (molecular weight). Larger proteins migrate faster through the column as they cannot penetrate the pores of the stationary phase as easily as smaller proteins.
  • Application: Particularly useful for the investigation of aggregates and oligomerisation states of proteins as well as for the determination of molecular weight.
  • Advantage: Native conditions, as there is no denaturation by solvents.
  • Disadvantage: Lower resolution compared to other methods.

 

3. Ion exchange chromatography (IEX)

  • Principle: In IEX, proteins are separated according to their charge. A protein binds to either an anionic or cationic stationary phase, depending on its isoelectric point (pI). Elution is achieved by gradually increasing the salt concentration in the mobile phase.
  • Application: Very useful for separating proteins with different charges, e.g. isoform proteins or charged impurities.
  • Advantage: High separation efficiency for proteins with minimal differences in pI.
  • Disadvantage: Sensitive to fluctuations in pH.

 

4. Hydrophobic interaction chromatography (HIC)

  • Principle: In HIC, proteins are separated based on their hydrophobic interaction with the stationary phase. The binding of the proteins takes place in the presence of high salt concentrations, which increase their hydrophobicity. Elution is achieved by gradually reducing the salt concentration.
  • Application: Particularly suitable for the separation of proteins with native hydrophobic regions without the use of organic solvents.
  • Advantage: Gentle separation of proteins under native conditions.
  • Disadvantage: Not so suitable for very hydrophilic proteins.

 

5. Affinity chromatography

  • Principle: This method is based on the specific binding between a protein and a ligand that is bound to the stationary phase. An example is His-tag affinity chromatography, in which histidine-labelled proteins bind to nickel or cobalt ions.
  • Application: Particularly useful for the targeted purification of a protein from complex mixtures, such as cell lysates.
  • Advantage: High selectivity and purity of the target protein.
  • Disadvantage: Relatively expensive and may require special pre-treatment of the protein (e.g. tagging).

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Manufacturer overview

AMT

HPLC - Reversed Phase:

  • HALO Protein C4
  • HALO Protein ES-C18
  • HALO Protein Diphenyl

GL Sciences

HPLC - Reversed Phase:

  • ProteoSil C18
  • ProteoSil C4
  • MonoSelect RP-mAb

 

HPLC - HILIC:

  • ProteoSil HILIC

 

HPLC - SEC:

  • ProteoSil SEC

 

HPLC - Mixed-Mode:

  • MonoSelect nPEC

Sample preparation - desalination and preconcentration:

  • MonoTip C18

 

Sample preparation - Desalination:

  • GL-Tip SDB
  • GL-Tip GC

 

Sample preparation - enrichment of phosphoproteins:

  • Titansphere
  • MonoTip TiO

 

Sample preparation - purification and enrichment:

  • MonoSpin Series

Imtakt

HPLC - Reversed Phase:

  • Intrada WP-RP
  • Cadenza CW-C18

 

UHPLC - Reversed Phase:

  • Presto FF-C18

Macherey-Nagel

HPLC - Reversed Phase:

  • Nucleodur 300 C4 ec, 5µm
  • Nucleosil 300 C8, 5µm
  • Nucleosil 300 C4, 5µm
  • Nucleodur 300 C18 ec, 5µm

 

HPLC - IEX:

  • Nucleogel SCX, 8µm
  • Nucleogel SAX, 8µm

Mitsubishi

HPLC - IEX:

  • XtalSpeed SP01
  • XtalSpeed DA01

 

HPLC - SEC:

  • MCI GEL CQP06, 10µm
  • MCI GEL CQP10, 10µm
  • MCI GEL CQP30, 10µm

Nacalai Tesque

HPLC - IEX:

  • Cosmogel IEX Type Q

 

HPLC - Reversed Phase:

  • Cosmosil C18-AR-300, 5µm
  • Protein-R, 5µm

 

HPLC - SEC:

  • Cosmosil Diol-II, 5µm

 

HPLC - HIC:

  • Cosmosil HIC, 5µm

Osaka Soda

HPLC - Reversed Phase:

  • Capcell Core WP, 2.7µm
  • Capcell PAK C18 SG300, 5µm
  • Proteonavi, 5µm

Sepax

HPLC - HIC:

  • Proteomix HIC

 

HPLC - IEX:

  • Proteomix IEX

 

HPLC - Reversed Phase:

  • Proteomix RP

 

HPLC - SEC:

  • SRT
  • SRT-C
  • Unix
  • Unix-C
  • Zenix
  • Zenix-C

Shodex

HPLC - Reversed Phase:

  • Silica C18M 4D
  • Asahipak ODP-50 6D
  • Asahipak C4P-50 4D

 

HPLC - SEC:

  • Protein KW Series
  • Protein LW Series
  • OHpak SB-802 HQ
  • Asahipak GF-510 HQ

HPLC - IEX:

  • IEC SP-FT
  • IEC QA-825
  • IEC SP-825
  • IEC DEAE-825
  • IEC CM-825
  • Asahipak ES-502C 7C
  • Asahipak ES-502N 7C

 

HPLC - Mixed-Mode:

  • Asahipak GS-320 7G

Thermo

HPLC - Reversed Phase:

  • Accucore 150-C4
  • ProSwift RP
  • PepSwift

 

HPLC - HIC:

  • MAbPAC HIC Series
  • ProPac HIC

 

HPLC - IEX:

  • ProPac Elite WCX
  • ProPac SAX-1
  • MAbPac SCX
  • ProPac 3R IEX

 

HPLC - metal affinity chromatography:

  • ProPac IMAC-10

Tosoh

HPLC - SEC:

  • TSKgel SW
  • TSKgel SWXL
  • TSKgel SuperSW
  • TSKgel UltraSW
  • TSKgel UP-SW

 

Proteins - IEX:

  • TSKgel BioAssist S
  • TSKgel BioAssist Q
  • TSKgel Q-STAT
  • TSKgel SP-5PW
  • TSKgel DEAE-5PW
  • TSKgel CM-5PW
  • TSKgel SP-STAT
  • TSKgel CM-STAT
  • TSKgel SP-NPR
  • TSKgel DEAE-NPR
  • TSKgel SuperQ-5PW

 

Proteins - HIC:

  • TSKgel Phenyl-5PW
  • TSKgel Ether-5PW
  • TSKgel Butyl-NPR

 

Proteins - Reversed Phase:

  • TSKgel Protein C4-300
  • TSKgel TMS-250

Waters

UHPLC - HILIC:

  • ACQUITY Glycoprotein BEH Amide, 1.7µm

 

UHPLC - HIC:

  • Protein-Pak Hi Res HIC, 2.5µm

 

HPLC - HIC:

  • Protein-Pak Hi Phenyl, 10µm

 

UHPLC - IEX:

  • BioSuite Anion-Exchange, 2.5µm

HPLC - IEX:

  • BioResolve SCX mAb, 3µm
  • BioSuite DEAE Anion-Exchange, 10µm
  • BioSuite SP Cation-Exchange, 10µm
  • BioSuite Cation-Exchange, 7µm
  • BioSuite CM Cation-Exchange, 10µm
  • BioSuite Q Anion-Exchange, 10µm
  • Protein-Pak Hi Res SP, 7µm
  • Protein-Pak Hi Res Q, 5µm
  • Protein-Pak DEAE 5PW, 12.5µm
  • Protein-Pak SP 5PW, 10µm
  • Protein-Pak Hi Res CM, 7µm

 

UHPLC - Reversed Phase:

  • ACQUITY Protein BEH C4, 1.7µm
  • nanoEase MZ Protein BEH C4, 1.7µm

HPLC - Reversed Phase:

  • BioResolve RP mAb Polyphenyl, 2.7µm
  • BioSuite Phenyl, 10µm
  • DeltaPak C4
  • Symmetry C4 300, 3.5µm, 5µm
  • XBridge Protein BEH C4

 

UHPLC - SEC:

  • ACQUITY Premier Protein SEC
  • BioResolve SEC mAb, 2.5µm
  • XBridge Premier Protein SEC, 2.5µm

 

HPLC - SEC:

  • BioSuite Diol (OH), 5µm
  • Protein-Pak SEC

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