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Time-saving PureCube Purification Technologies 

Benefit from affinity beads developed by experts with 30 years of experience

Recombinant protein production | Membrane protein purification | Protein-protein interaction studies | Co-immunoprecipitation | Mass spectrometry sample preparation | ELISA | Western Blot detection | Protein localization |

Technologies: His-tag Ni-INDIGO | Ni-NTA | Rho1D4-tag | Strep-tag | Surface groups
Protein research is driven by our shared passion for discovery and methodological improvement. During protein purification our customers often report challenges such as high levels of unspecific binding, aggregated protein, nickel-ion leakage, the need for specific customized resin, low yield and purity. These challenges drive us to develop innovative products that transcend conventional protein purification methods, enabling customers to make valuable contributions to our field.

Technology 1: His-tag Affinity

Highest protein yields due to Ni-Leakage resistant beads

PureCube Ni-INDIGO Agarose and MagBeads

PureCube Ni-INDIGO technology offers you:

  • Up to 30 times lower Ni Leakage
  • Yields of 80-100 mg/ml resin (See Fig. 1)
  • Higher protein purity compared to competitor Ni-NTA products (See Fig. 2)
  • Resistance to EDTA, DTT, Phenanthroline, GuHCl, TCEP, Triton, Urea, Acetonitrile, etc.
  • 50% reduction in the regeneration process
  • Reusability with consistent binding capacity

Figure 1: Yield of recoverd GFP in a purification batch process with CubePure Ni-INDIGO Agarose with different tested additives 0.1M Citrate, 20 mM DTT, 6 mM GuHCl, 8 M GuHCl, 10 TCEP, 2% Titron, 8 M Urea, 20 mM EDTA.

Figure 2: Comparison of the protein purity of a His-tag purification assay. PureCube Ni-INDIGO agarose from Cube Biotech in contrast to Ni-NTA agarose from market competitor "T". Both purifications were performed under the same conditions.

Figure 3: Protein yield of His-tagged protein compared between PureCube Ni-INDIGO based beads and competitor products at 20 mM EDTA and 10 mM DTT.

PureCube Ni-INDIGO-Technology in research

Michael Crone, PhD,
Solena Materials,
London

"The combination of high protein binding capacity and ease of use makes the PureCube Ni-INDIGO resin our go-to product when it comes to Ni-affinity purification.

The resin seems to retain its full protein binding capacity even after 30 uses when each purification is followed by the recommended CIP protocol. The INDIGO technology allows you to clean the resin with 0.5 M NaOH without stripping the Nickel thus eliminating tedious resin metal ion stripping and recharging. We would highly recommend PureCube Ni-INDIGO resin to anyone doing Ni-Affinity protein purification."
PureCube Ni-INDIGO was utilized to obtain high-purity Mpro, which is necessary for testing the efficacy of the newly designed inhibitors. "High purity protein was eluted in a buffer of 50 mM Tris (pH 8), 250 mM Imidazole, and 300 mM NaCl. Eluted fractions with high protein content were pooled and buffer exchanged into HRV protease cleavage buffer..."

Westberg, et al. 2020
The effectiveness of the PureCube Ni-INDIGO column was utilized for the purification of ACE2, facilitating subsequent investigation of binding interactions via microarray assays. "We isolated seven new multiformin azaphilones and assigned the stereochemistry of all stereocenters"

Jansen-Olliges, et al. 2022

Technology 2: Ni-NTA Affinity

Evolution of Ni-NTA: Beads with the highest ligand density

Ni-NTA Agarose and MagBeads

PureCube Ni-NTA beads with highest ligand density enable you to:

  • produce protein yields that outperform all leading competitor materials (Fig. 4 & 5)
  • maintain over 20% higher protein binding capacity

Figure 4: His-tagged GFP was purified from 1. E.coli cell lysates, and 2. Insect cell lysates on His affinity resins in batch format. PureCube Ni-NTA Agarose was compared to conventional Ni-NTA resins of leading competitors, we refer to them as “A“, “B“, and “C“. Recovered GFP was quantified by absorption at 488 nm. ​Binding E.coli lysates: ​50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, 1 mM EDTA, 10 mM DTT, pH 7.4​; Binding Insect cell supernatants: Sample as is with 10 mM imidazole added​; Wash: 50 mM NaH2PO4, 300 mM NaCl, 20 mM imidazole, 1 mM EDTA, 10 mM DTT, pH 7.4​; Elution: 50 mM NaH2PO4, 300 mM NaCl, 250 mM imidazole, 1 mM EDTA, 10 mM DTT, pH 7.4

Figure 5: Over 20% more yield obtained with PureCube Ni-NTA Agarose. SDS-PAGE of GFP expressed in E. coli and purified in gravity columns with PureCube Ni-NTA Agarose and Ni-NTA resin from Competitor G, and Competitor Q. 80 mg/mL protein yield was obtained with PureCube Ni-NTA Agarose (E1–E4, Cube) compared to 65 and 48 mg/mL, respectively, with the widely used alternative resins G and Q (E1–E4, Competitor G / Competitor Q).

Technology 3: Rho1D4-tag Affinity

Unmatched purity for low-abundance proteins

HighSpec Rho1D4 Agarose

  • Ideal for membrane proteins
  • Highest specificity (KD = 20 nM)
  • One-step purification
  • Mild elution conditions
  • Rho1D4 monoclonal antibodies minimize non-specific bindings
  • Perfect for applications needing small and pure sample amounts, e.g. Cryo-EM

Fig. 6: Purification of chemokine receptor 4 (CXCR4) using PureCube Rho1D4 Agarose. Total lysate (TL) was solubilized with Fos-Choline-14, and the soluble fraction (SF) was incubated on an immunoaffinity column loaded with rho1D4 antibody. The concentration of eluted CXCR4 ranged 0.6-1.0 mg/mL as determined spectrophotometrically.

"My working group uses rho1D4 resins from Cube Biotech to purify membrane proteins. For our applications we need very high purity. The rho1D4-tag provides high specificity  and yield at the same time. (...) We will certainly use it again in the future! "

Prof. Dr. Jörg Labahn, Group leader at Centre for Structural Systems Biology

Technology 4:  Strep-tag® Affinity

The best balance of specificity and purity

StrepTactin® Agarose and Magbeads

  • Highly selective binding properties leading to unparalleled protein purity (>95%) 
  • Exceptionally high affinity in a picomolar range to Twin-Strep-tag® enables reliable use in analytic applications
  • Protein yields of up to 31 mg/ml agarose resin and 42.5 mg/ml magnetic beads 
  • Mild elution conditions that preserve natural protein structure 
  •  Variability in buffer conditions, e.g. high salts, detergents, metal ions, chelators or reducing agents

Figure 7: Strep-Tactin®XT 4Flow high capacity binding capacity during purification of CD31 fab fragment (50.3 kDa) and CD56 antigen (82.6 kDa ) fused to Twin-Strep-tag®.

Figure 8: CD31 fab fragment fused to Twin-Strep-tag® was isolated from E. coli lysate with Strep-Tactin®XT 4Flow® high capacity with a purity of 95%.

Technology 5: Active surface groups

Unimagined possibilities with customized beads

Activated Resins & MagBeads

  1. Amine activated beads 
  2. Carboxy activated beads 
  3. Epoxy activated beads 
  4.  Maleimide activated beads 
  5. NHS activated beads
  • Bind a vast variety of proteins e.g. antibody fragments, peptides or small molecules
  • Stable bond due to covalent binding
  • Use for immunoprecipitation (Pull-down assay), affinity purification, diagnostics (ELISA), drug delivery and biosensors

CubePure Epoxy Activated Beads in research

Video of the Intelligent Infusion Masterclass: Thoman Bortecen about CubePure Epoxy Activated Beads using it for s semi-automated workflow for the quantitative analysis of the newly synthesized proteome.

Through the use of novel magnetic alkyne beads, the enrichment of newly synthesized proteins can be performed with a semi-automated protocol using 10-fold lower input.

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Reach out to us

Anton Starke
Strategic Partnership Manager
Protein Purification
Anton.Starke@cube-biotech.com
+49-2173-99373-16

Sebastian Lühn
Strategic Partnership Manager
Protein Purification
Anton.Starke@cube-biotech.com
+49-2173-99373-16